Novel formulation of naproxen

ABSTRACT

The present invention relates to methods for producing particles of naproxen using dry milling processes as well as compositions comprising naproxen, medicaments produced using naproxen in particulate form and/or compositions, and to methods of treatment of an animal, including man, using a therapeutically effective amount of naproxen administered by way of said medicaments.

RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.13/266,119, filed Feb. 15, 2012, which is a continuation ofInternational Application Number PCT/AU2010/000470, filed on 23 Apr.2010, which claims priority to AU Application No. 2009901746, filed on24 Apr. 2009 and U.S. Application No. 61/172,289, filed on 24 Apr. 2009,the entire contents of which applications are hereby incorporated byreference.

FIELD OF THE INVENTION

The present invention relates to methods for producing particles ofnaproxen using dry milling processes as well as compositions comprisingnaproxen, medicaments produced using naproxen in particulate form and/orcompositions, and to methods of treatment of an animal, including man,using a therapeutically effective amount of naproxen administered by wayof said medicaments.

BACKGROUND

Poor bioavailability is a significant problem encountered in thedevelopment of compositions in the therapeutic, cosmetic, agriculturaland food industries, particularly those materials containing abiologically active material that is poorly soluble in water atphysiological pH. An active material's bioavailability is the degree towhich the active material becomes available to the target tissue in thebody or other medium after systemic administration through, for example,oral or intravenous means. Many factors affect bioavailability,including the form of dosage and the solubility and dissolution rate ofthe active material.

In therapeutic applications, poorly and slowly water-soluble materialstend to be eliminated from the gastrointestinal tract before beingabsorbed into the circulation. In addition, poorly soluble active agentstend to be disfavored or even unsafe for intravenous administration dueto the risk of particles of agent blocking blood flow throughcapillaries.

It is known that the rate of dissolution of a particulate drug willincrease with increasing surface area. One way of increasing surfacearea is decreasing particle size. Consequently, methods of making finelydivided or sized drugs have been studied with a view to controlling thesize and size range of drug particles for pharmaceutical compositions.

For example, dry milling techniques have been used to reduce particlesize and hence influence drug absorption. However, in conventional drymilling the limit of fineness is reached generally in the region ofabout 100 microns (100,000 nm), at which point material cakes on themilling chamber and prevents any further diminution of particle size.Alternatively, wet grinding may be employed to reduce particle size, butflocculation restricts the lower particle size limit to approximately 10microns (10,000 nm). The wet milling process, however, is prone tocontamination, thereby leading to a bias in the pharmaceutical artagainst wet milling. Another alternative milling technique, commercialairjet milling, has provided particles ranging in average size from aslow as about 1 to about 50 microns (1,000-50,000 nm).

There are several approaches currently used to formulate poorly solubleactive agents. One approach is to prepare the active agent as a solublesalt. Where this approach cannot be employed, alternate (usuallyphysical) approaches are employed to improve the solubility of theactive agent. Alternate approaches generally subject the active agent tophysical conditions that change the agent's physical and or chemicalproperties to improve its solubility. These include process technologiessuch as micronization, modification of crystal or polymorphic structure,development of oil based solutions, use of co-solvents, surfacestabilizers or complexing agents, micro-emulsions, super critical fluidand production of solid dispersions or solutions. More than one of theseprocesses may be used in combination to improve formulation of aparticular therapeutic material. Many of these approaches commonlyconvert a drug into an amorphous state, which generally leads to ahigher dissolution rate. However, formulation approaches that result inthe production of amorphous material are not common in commercialformulations due to concerns relating to stability and the potential formaterial to re-crystallize.

These techniques for preparing such pharmaceutical compositions tend tobe complex. By way of example, a principal technical difficultyencountered with emulsion polymerization is the removal of contaminants,such as unreacted monomers or initiators (which may have undesirablelevels of toxicity), at the end of the manufacturing process.

Another method of providing reduced particle size is the formation ofpharmaceutical drug microcapsules, which techniques include micronizing,polymerisation and co-dispersion. However, these techniques suffer froma number of disadvantages including at least the inability to producesufficiently small particles such as those obtained by milling, and thepresence of co-solvents and/or contaminants such as toxic monomers whichare difficult to remove, leading to expensive manufacturing processes.

Over the last decade, intense scientific investigation has been carriedout to improve the solubility of active agents by converting the agentsto ultra fine powders by methods such as milling and grinding. Thesetechniques may be used to increase the dissolution rate of a particulatesolid by increasing the overall surface area and decreasing the meanparticle size. U.S. Pat. No. 6,634,576 discloses examples of wet-millinga solid substrate, such as a pharmaceutically active compound, toproduce a “synergetic co-mixture”.

International Patent Application PCT/AU2005/001977 (NanoparticleComposition(s) and Method for Synthesis Thereof) describes, inter alia,a method comprising the step of contacting a precursor compound with aco-reactant under mechanochemical synthesis conditions wherein asolid-state chemical reaction between the precursor compound and theco-reactant produces therapeutically active nanoparticles dispersed in acarrier matrix. Mechanochemical synthesis, as discussed in InternationalPatent Application PCT/AU2005/001977, refers to the use of mechanicalenergy to activate, initiate or promote a chemical reaction, a crystalstructure transformation or a phase change in a material or a mixture ofmaterials, for example by agitating a reaction mixture in the presenceof a milling media to transfer mechanical energy to the reactionmixture, and includes without limitation “mechanochemical activation”,“mechanochemical processing”, “reactive milling”, and related processes.

International Patent Application PCT/AU2007/000910 (Methods for thepreparation of biologically active compounds in nanoparticulate form)describes, inter alia, a method for dry milling raloxifene with lactoseand NaCl which produced nanoparticulate raloxifene without significantaggregation problems. The methods disclosed by the prior art producenanoparticles at volume fractions of 15% or less and suggests that 25%is the upper limit for the volume fraction of the biologically activematerial that could be successfully converted to smaller particles.

The present invention provides methods for an improved milling processwhich produces particles of active compound with increased surface area,yet allows for higher volume fractions of the biologically activematerial.

One example of a therapeutic area where this technology could be appliedin is the area of acute pain management. Many pain medications such asnaproxen provides pain relief for chronic pain. As a result they arecommonly taken on a daily basis to maintain an effective therapeuticlevel. Because naproxen is a poorly water soluble drug dissolution andabsorbtion to the body is slow with the Tmax of current commercialformulations in the range of 1-4 hours. So a method such as the presentinvention which provides for improved dissolution, will likely providemuch faster absorption resulting in a more rapid onset of thetherapeutic effect. By using a method such as the present invention,which provides faster absorption, a drug such as naproxen, could be usedmore readily to treat acute pain as well as chronic pain.

Naproxen dosages typically range from 200-500 mg of active. Because ofthis requirement for high amounts of active ingredient pervious artwhich produced nanoparticles at 15% would be difficult to use to producea commercial formulation. As the present invention provides for theproduction of particles at higher volume fractions is it more suitablefor medications such as naproxen.

Although the background to the present invention is discussed in thecontext of improving the bioavailability of materials that are poorly orslowly water soluble, the applications of the methods of the presentinvention are not limited to such, as is evident from the followingdescription of the invention.

Further, although the background to the present invention is largelydiscussed in the context of improving the bioavailability of therapeuticor pharmaceutical compounds, the applications of the methods of thepresent invention are clearly not limited to such. For example, as isevident from the following description, applications of the methods ofthe present invention include but are not limited to: nutraceutical andnutritional compounds, complementary medicinal compounds, veterinarytherapeutic applications and agricultural chemical applications, such aspesticide, fungicide or herbicide.

Furthermore an application of the current invention would be tomaterials which contain a biologically active compound such as, but notlimited to a therapeutic or pharmaceutical compound, a nutraceutical ornutrient, a complementary medicinal product such as active components inplant or other naturally occurring material, a veterinary therapeuticcompound or an agricultural compound such as a pesticide, fungicide orherbicide. Specific examples would be the spice turmeric that containsthe active compound curcumin, or flax seed that contains the nutrientALA an omega 3 fatty acid. As these specific examples indicate thisinvention could be applied to, but not limited to, a range of naturalproducts such as seeds, cocoa and cocoa solids, coffee, herbs, spices,other plant materials or food materials that contain a biologicallyactive compound. The application of this invention to these types ofmaterials would enable greater availability of the active compound inthe materials when used in the relevant application. For example wherematerial subject to this invention is orally ingested the active wouldbe more bioavailable.

SUMMARY OF THE INVENTION

In one aspect the present invention is directed to the unexpectedfinding that particles of a biologically active material can be producedby dry milling processes wherein the composition produced by said methodcomprises particles of the biologically active material at or above avolume fraction of 25 v/v %. In one surprising aspect the particle sizeproduced by the process is equal to or less than 2000 nm. In anothersurprising aspect the particle size produced by the process is equal toor less than 1000 nm. In another surprising aspect the crystallinity ofthe active material is unchanged or not substantially changed. In apreferred embodiment the present invention is directed to the unexpectedfinding that particles of naproxen can be produced by dry millingprocesses at commercial scale.

Preferably the method comprises particles of the biologically activematerial at or above a volume fraction selected from the groupconsisting of 25 v/v %; 30 v/v %; 35 v/v %; 40 v/v %; 45 v/v %; 50 v/v%, 55 v/v % and 60 v/v %. Preferably the method comprises particles ofthe biologically active material at or below a volume fraction selectedfrom the group consisting of 60 v/v %, 55 v/v %, 50 v/v %; 45 v/v %; 40v/v %; and 35 v/v %.

Thus in a first aspect the invention comprises a method producing acomposition, comprising the steps of dry milling a solid biologicallyactive material and a millable grinding matrix in a mill comprising aplurality of milling bodies, for a time period sufficient to produceparticles of the biologically active material dispersed in an at leastpartially milled grinding material, wherein the composition produced bysaid method comprises particles of the biologically active material ator above a volume fraction of 25 v/v %.

In one preferred embodiment, the average particle size, determined on aparticle number basis, is equal to or less than a size selected from thegroup 2000 nm, 1900 nm, 1800 nm, 1700 nm, 1600 nm, 1500 nm, 1400 nm,1300 nm, 1200 nm, 1100 nm, 1000 nm, 900 nm, 800 nm, 700 nm, 600 nm, 500nm, 400 nm, 300 nm, 200 nm and 100 nm. Preferably, the average particlesize is equal to or greater than 25 nm.

In another preferred embodiment, the particles have a median particlesize, determined on a particle volume basis, equal or less than a sizeselected from the group 2000 nm, 1900 nm, 1800 nm, 1700 nm, 1600 nm,1500 nm, 1400 nm, 1300 nm, 1200 nm, 1100 nm, 1000 nm, 900 nm, 800 nm,700 nm, 600 nm, 500 nm, 400 nm, 300 nm, 200 nm and 100 nm. Preferably,the median particle size is equal to or greater than 25 nm. Preferably,the percentage of particles, on a particle volume basis, is selectedfrom the group consisting of: 50%, 60%, 70%, 80%, 90%, 95% and 100% lessthan 2000 nm (%<2000 nm). Preferably, the percentage of particles, on aparticle volume basis, is selected from the group consisting of: 50%,60%, 70%, 80%, 90%, 95% and 100% less than 1000 nm (%<1000 nm).Preferably, the percentage of particles, on a particle volume basis, isselected from the group 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%,95% and 100% less than 500 nm (%<500 nm). Preferably, the percentage ofparticles, on a particle volume basis, is selected from the group 0%,10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% and 100% less than 300nm (%<300 nm). Preferably, the percentage of particles, on a particlevolume basis, is selected from the group 0%, 10%, 20%, 30%, 40%, 50%,60%, 70%, 80%, 90%, 95% and 100% less than 200 nm (%<200 nm).Preferably, the Dx of the particle size distribution, as measured on aparticle volume basis, is selected from the group consisting of lessthan or equal to 10,000 nm, 5000 nm, 3000 nm, 2000 nm, 1900 nm, 1800 nm,1700 nm, 1600 nm, 1500 nm, 1400 nm, 1300 nm, 1200 nm, 1100 nm, 1000 nm,900 nm, 800 nm, 700 nm, 600 nm, 500 nm, 400 nm, 300 nm, 200 nm, and 100nm; wherein x is greater than or equal to 90.

In another preferred embodiment, the crystallinity profile of thebiologically active material is selected from the group consisting of:at least 50% of the biologically active material is crystalline, atleast 60% of the biologically active material is crystalline, at least70% of the biologically active material is crystalline, at least 75% ofthe biologically active material is crystalline, at least 85% of thebiologically active material is crystalline, at least 90% of thebiologically active material is crystalline, at least 95% of thebiologically active material is crystalline and at least 98% of thebiologically active material is crystalline. More preferably, thecrystallinity profile of the biologically active material issubstantially equal to the crystallinity profile of the biologicallyactive material before the material was subjected to the method asdescribed herein.

In another preferred embodiment, the amorphous content of thebiologically active material is selected from the group consisting of:less than 50% of the biologically active material is amorphous, lessthan 40% of the biologically active material is amorphous, less than 30%of the biologically active material is amorphous, less than 25% of thebiologically active material is amorphous, less than 15% of thebiologically active material is amorphous, less than 10% of thebiologically active material is amorphous, less than 5% of thebiologically active material is amorphous and less than 2% of thebiologically active material is amorphous. Preferably, the biologicallyactive material has no significant increase in amorphous content aftersubjecting the material to the method as described herein.

In another preferred embodiment, the milling time period is a rangeselected from the group consisting of: between 10 minutes and 2 hours,between 10 minutes and 90 minutes, between 10 minutes and 1 hour,between 10 minutes and 45 minutes, between 10 minutes and 30 minutes,between 5 minutes and 30 minutes, between 5 minutes and 20 minutes,between 2 minutes and 10 minutes, between 2 minutes and 5 minutes,between 1 minutes and 20 minutes, between 1 minute and 10 minutes, andbetween 1 minute and 5 minutes.

In another preferred embodiment, the milling medium is selected from thegroup consisting of: ceramics, glasses, polymers, ferromagnetics andmetals. Preferably, the milling medium is steel balls having a diameterselected from the group consisting of: between 1 and 20 mm, between 2and 15 mm and between 3 and 10 mm. In another preferred embodiment, themilling medium is zirconium oxide balls having a diameter selected fromthe group consisting of: between 1 and 20 mm, between 2 and 15 mm andbetween 3 and 10 mm. Preferably, the dry milling apparatus is a millselected from the group consisting of: attritor mills (horizontal orvertical), nutating mills, tower mills, pearl mills, planetary mills,vibratory mills, eccentric vibratory mills, gravity-dependent-type ballmills, rod mills, roller mills and crusher mills. Preferably, themilling medium within the milling apparatus is mechanically agitated by1, 2 or 3 rotating shafts. Preferably, the method is configured toproduce the biologically active material in a continuous fashion.

Preferably, the total combined amount of biologically active materialand grinding matrix in the mill at any given time is equal to or greaterthan a mass selected from the group consisting of: 200 grams, 500 grams,1 kg, 2 kg, 5 kg, 10 kg, 20 kg, 30 kg, 50 kg, 75 kg, 100 kg, 150 kg, 200kg. Preferably, the total combined amount of biologically activematerial and grinding matrix is less than 2000 kg.

In another preferred embodiment, the grinding matrix is a singlematerial or is a mixture of two or more materials in any proportion.Preferably, the single material or a mixture of two or more materials isselected from the group consisting of: mannitol, sorbitol, Isomalt,xylitol, maltitol, lactitol, erythritol, arabitol, ribitol, glucose,fructose, mannose, galactose, anhydrous lactose, lactose monohydrate,sucrose, maltose, trehalose, maltodextrins, dextrin, Inulin, dextrates,polydextrose, starch, wheat flour, corn flour, rice flour, rice starch,tapioca flour, tapioca starch, potato flour, potato starch, other floursand starches, milk powder, skim milk powders, other milk solids anddreviatives, soy flour, soy meal or other soy products, cellulose,microcystalline cellulose, microcystalline cellulose based co blendedmaterials, pregelatinized (or partially) starch, HPMC, CMC, HPC, citricacid, tartaric acid, malic acid, maleic acid fumaric acid, ascorbicacid, succinic acid, sodium citrate, sodium tartrate, sodium malate,sodium ascorbate, potassium citrate, potassium tartrate, potassiummalate, potassium ascorbate, sodium carbonate, potassium carbonate,magnesium carbonate, sodium bicarbonate, potassium bicarbonate andcalcium carbonate. dibasic calcium phosphate, tribasic calciumphosphate, sodium sulfate, sodium chloride, sodium metabisulphite,sodium thiosulfate, ammonium chloride, Glauber's salt, ammoniumcarbonate, sodium bisulfate, magnesium sulfate, potash alum, potassiumchloride, sodium hydrogen sulfate, sodium hydroxide, crystallinehydroxides, hydrogen carbonates, ammonium chloride, methylaminehydrochloride, ammonium bromide, silica, thermal silica, alumina,titanium dioxide, talc, chalk, mica, kaolin, bentonite, hectorite,magnesium trisilicate, clay based materials or aluminium silicates,sodium lauryl sulfate, sodium stearyl sulfate, sodium cetyl sulfate,sodium cetostearyl sulfate, sodium docusate, sodium deoxycholate,N-lauroylsarcosine sodium salt, glyceryl monostearate, glyceroldistearate glyceryl palmitostearate, glyceryl behenate, glycerylcaprylate, glyceryl oleate, benzalkonium chloride, CTAB, CTAC,Cetrimide, cetylpyridinium chloride, cetylpyridinium bromide,benzethonium chloride, PEG 40 stearate, PEG 100 stearate, poloxamer 188,338, poloxamer 407 polyoxyl 2 stearyl ether, polyoxyl 100 stearyl ether,polyoxyl 20 stearyl ether, polyoxyl 10 stearyl ether, polyoxyl 20 cetylether, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 61,polysorbate 65, polysorbate 80, polyoxyl 35 castor oil, polyoxyl 40castor oil, polyoxyl 60 castor oil, polyoxyl 100 castor oil, polyoxyl200 castor oil, polyoxyl 40 hydrogenated castor oil, polyoxyl 60hydrogenated castor oil, polyoxyl 100 hydrogenated castor oil, polyoxyl200 hydrogenated castor oil, cetostearyl alcohol, macrogel 15hydroxystearate, sorbitan monopalmitate, sorbitan monostearate, sorbitantrioleate, Sucrose Palmitate, Sucrose Stearate, Sucrose Distearate,Sucrose laurate, Glycocholic acid, sodium Glycholate, Cholic Acid,Sodium Cholate, Sodium Deoxycholate, Deoxycholic acid, Sodiumtaurocholate, taurocholic acid, Sodium taurodeoxycholate,taurodeoxycholic acid, soy lecithin, phosphatidylcholine,phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol,PEG4000, PEG6000, PEG8000, PEG10000, PEG20000, alkyl naphthalenesulfonate condensate/Lignosulfonate blend, Calcium DodecylbenzeneSulfonate, Sodium Dodecylbenzene Sulfonate, Diisopropylnaphthaenesulphonate, erythritol distearate, Naphthalene SulfonateFormaldehyde Condensate, nonylphenol ethoxylate (poe-30),Tristyrylphenol Ethoxylate, Polyoxyethylene (15) tallowalkylamines,sodium alkyl naphthalene sulfonate, sodium alkyl naphthalene sulfonatecondensate, sodium alkylbenzene sulfonate, sodium isopropyl naphthalenesulfonate, Sodium Methyl Naphthalene Formaldehyde Sulfonate, sodiumn-butyl naphthalene sulfonate, tridecyl alcohol ethoxylate (poe-18),Triethanolamine isodecanol phosphate ester, Triethanolaminetristyrylphosphate ester, Tristyrylphenol Ethoxylate Sulfate,Bis(2-hydroxyethyl)tallowalkylamines. Preferably, the concentration ofthe single (or first) material is selected from the group consisting of:5-99% w/w, 10-95% w/w, 15-85% w/w, of 20-80% w/w, 25-75% w/w, 30-60%w/w, 40-50% w/w. Preferably, the concentration of the second orsubsequent material is selected from the group consisting of: 5-50% w/w,5-40% w/w, 5-30% w/w, of 5-20% w/w, 10-40% w/w, 10-30% w/w, 10-20% w/w,20-40% w/w, or 20-30% w/w or if the second or subsequent material is asurfactant or water soluble polymer the concentration is selected from0.1-10% w/w, 0.1-5% w/w, 0.1-2.5% w/w, of 0.1-2% w/w, 0.1-1%, 0.5-5%w/w, 0.5-3% w/w, 0.5-2% w/w, 0.5-1.5%, 0.5-1% w/w, of 0.75-1.25% w/w,0.75-1% and 1% w/w.

Preferably, the grinding matrix is selected from the group consistingof:

-   -   (a) lactose monohydrate or lactose monohydrate combined with at        least one material selected from the group consisting of:        xylitol; lactose anhydrous; microcrystalline cellulose; sucrose;        glucose; sodium chloride; talc; kaolin; calcium carbonate; malic        acid; trisodium citrate dihydrate; D,L-Malic acid; sodium        pentane sulfate; sodium octadecyl sulfate; Brij700; Brij76;        sodium n-lauroyl sacrosine; lecithin; docusate sodium;        polyoxyl-40-stearate; Aerosil R972 fumed silica; sodium lauryl        sulfate or other alkyl sulfate surfactants with a chain length        between C5 to C18; polyvinyl pyrrolidone; sodium lauryl sulfate        and polyethylene glycol 40 stearate, sodium lauryl sulfate and        polyethylene glycol 100 stearate, sodium lauryl sulfate and PEG        3000, sodium lauryl sulphate and PEG 6000, sodium lauryl        sulphate and PEG 8000, sodium lauryl sulphate and PEG 10000,        sodium lauryl sulfate and Brij700, sodium lauryl sulfate and        Poloxamer 407, sodium lauryl sulfate and Poloxamer 338, sodium        lauryl sulfate and Poloxamer 188; Poloxamer 407, Poloxamer 338,        Poloxamer 188, alkyl naphthalene sulfonate        condensate/Lignosulfonate blend; Calcium Dodecylbenzene        Sulfonate (Branched); Diisopropyl naphthalenesulphonate;        erythritol distearate; linear and branched dodecylbenzene        sulfonic acids; Naphthalene Sulfonate Formaldehyde Condensate;        nonylphenol ethoxylate, POE-30; Phosphate Esters,        Tristyrylphenol Ethoxylate, Free Acid; Polyoxyethylene (15)        tallowalkylamines; sodium alkyl naphthalene sulfonate; sodium        alkyl naphthalene sulfonate condensate; sodium alkylbenzene        sulfonate; sodium isopropyl naphthalene sulfonate; Sodium Methyl        Naphthalene; Formaldehyde Sulfonate; sodium salt of n-butyl        naphthalene sulfonate; tridecyl alcohol ethoxylate, POE-18;        Triethanolamine isodecanol phosphate ester; Triethanolamine        tristyrylphosphate ester; Tristyrylphenol Ethoxylate Sulfate;        Bis(2-hydroxyethyl)tallowalkylamines.    -   (b) lactose anhydrous or lactose anhydrous combined with at        least one material selected from the group consisting of:        lactose monohydrate; xylitol; microcrystalline cellulose;        sucrose; glucose; sodium chloride; talc; kaolin; calcium        carbonate; malic acid; trisodium citrate dihydrate; D,L-Malic        acid; sodium pentane sulfate; sodium octadecyl sulfate; Brij700;        Brij76; sodium n-lauroyl sacrosine; lecithin; docusate sodium;        polyoxyl-40-stearate; Aerosil R972 fumed silica; sodium lauryl        sulfate or other alkyl sulfate surfactants with a chain length        between C5 to C18; polyvinyl pyrrolidone; sodium lauryl sulfate        and polyethylene glycol 40 stearate, sodium lauryl sulfate and        polyethylene glycol 100 stearate, sodium lauryl sulfate and PEG        3000, sodium lauryl sulphate and PEG 6000, sodium lauryl        sulphate and PEG 8000, sodium lauryl sulphate and PEG 10000,        sodium lauryl sulfate and Brij700, sodium lauryl sulfate and        Poloxamer 407, sodium lauryl sulfate and Poloxamer 338, sodium        lauryl sulfate and Poloxamer 188; Poloxamer 407, Poloxamer 338,        Poloxamer 188, alkyl naphthalene sulfonate        condensate/Lignosulfonate blend; Calcium Dodecylbenzene        Sulfonate (Branched); Diisopropyl naphthalenesulphonate;        erythritol distearate; linear and branched dodecylbenzene        sulfonic acids; Naphthalene Sulfonate Formaldehyde Condensate;        nonylphenol ethoxylate, POE-30; Phosphate Esters,        Tristyrylphenol Ethoxylate, Free Acid; Polyoxyethylene (15)        tallowalkylamines; sodium alkyl naphthalene sulfonate; sodium        alkyl naphthalene sulfonate condensate; sodium alkylbenzene        sulfonate; sodium isopropyl naphthalene sulfonate; Sodium Methyl        Naphthalene; Formaldehyde Sulfonate; sodium salt of n-butyl        naphthalene sulfonate; tridecyl alcohol ethoxylate, POE-18;        Triethanolamine isodecanol phosphate ester; Triethanolamine        tristyrylphosphate ester; Tristyrylphenol Ethoxylate Sulfate;        Bis(2-hydroxyethyl)tallowalkylamines.    -   (c) mannitol or mannitol combined with at least one material        selected from the group consisting of: lactose monohydrate;        xylitol; lactose anhydrous; microcrystalline cellulose; sucrose;        glucose; sodium chloride; talc; kaolin; calcium carbonate; malic        acid; trisodium citrate dihydrate; D,L-Malic acid; sodium        pentane sulfate; sodium octadecyl sulfate; Brij700; Brij76;        sodium n-lauroyl sacrosine; lecithin; docusate sodium;        polyoxyl-40-stearate; Aerosil R972 fumed silica; sodium lauryl        sulfate or other alkyl sulfate surfactants with a chain length        between C5 to C18; polyvinyl pyrrolidone; sodium lauryl sulfate        and polyethylene glycol 40 stearate, sodium lauryl sulfate and        polyethylene glycol 100 stearate, sodium lauryl sulfate and PEG        3000, sodium lauryl sulphate and PEG 6000, sodium lauryl        sulphate and PEG 8000, sodium lauryl sulphate and PEG 10000,        sodium lauryl sulfate and Brij700, sodium lauryl sulfate and        Poloxamer 407, sodium lauryl sulfate and Poloxamer 338, sodium        lauryl sulfate and Poloxamer 188; Poloxamer 407, Poloxamer 338,        Poloxamer 188, alkyl naphthalene sulfonate        condensate/Lignosulfonate blend; Calcium Dodecylbenzene        Sulfonate (Branched); Diisopropyl naphthalenesulphonate;        erythritol distearate; linear and branched dodecylbenzene        sulfonic acids; Naphthalene Sulfonate Formaldehyde Condensate;        nonylphenol ethoxylate, POE-30; Phosphate Esters,        Tristyrylphenol Ethoxylate, Free Acid; Polyoxyethylene (15)        tallowalkylamines; sodium alkyl naphthalene sulfonate; sodium        alkyl naphthalene sulfonate condensate; sodium alkylbenzene        sulfonate; sodium isopropyl naphthalene sulfonate; Sodium Methyl        Naphthalene; Formaldehyde Sulfonate; sodium salt of n-butyl        naphthalene sulfonate; tridecyl alcohol ethoxylate, POE-18;        Triethanolamine isodecanol phosphate ester; Triethanolamine        tristyrylphosphate ester; Tristyrylphenol Ethoxylate Sulfate;        Bis(2-hydroxyethyl)tallowalkylamines.    -   (d) Sucrose or sucrose combined with at least one material        selected from the group consisting of: lactose monohydrate;        lactose anhydrous; mannitol; microcrystalline cellulose;        glucose; sodium chloride; talc; kaolin; calcium carbonate; malic        acid; tartaric acid; trisodium citrate dihydrate; D,L-Malic        acid; sodium pentane sulfate; sodium octadecyl sulfate; Brij700;        Brij76; sodium n-lauroyl sacrosine; lecithin; docusate sodium;        polyoxyl-40-stearate; Aerosil R972 fumed silica; sodium lauryl        sulfate or other alkyl sulfate surfactants with a chain length        between C5 to C18; polyvinyl pyrrolidone; sodium lauryl sulfate        and polyethylene glycol 40 stearate, sodium lauryl sulfate and        polyethylene glycol 100 stearate, sodium lauryl sulfate and PEG        3000, sodium lauryl sulphate and PEG 6000, sodium lauryl        sulphate and PEG 8000, sodium lauryl sulphate and PEG 10000,        sodium lauryl sulfate and Brij700, sodium lauryl sulfate and        Poloxamer 407, sodium lauryl sulfate and Poloxamer 338, sodium        lauryl sulfate and Poloxamer 188; Poloxamer 407, Poloxamer 338,        Poloxamer 188, alkyl naphthalene sulfonate        condensate/Lignosulfonate blend; Calcium Dodecylbenzene        Sulfonate (Branched); Diisopropyl naphthalenesulphonate;        erythritol distearate; linear and branched dodecylbenzene        sulfonic acids; Naphthalene Sulfonate Formaldehyde Condensate;        nonylphenol ethoxylate, POE-30; Phosphate Esters,        Tristyrylphenol Ethoxylate, Free Acid; Polyoxyethylene (15)        tallowalkylamines; sodium alkyl naphthalene sulfonate; sodium        alkyl naphthalene sulfonate condensate; sodium alkylbenzene        sulfonate; sodium isopropyl naphthalene sulfonate; Sodium Methyl        Naphthalene; Formaldehyde Sulfonate; sodium salt of n-butyl        naphthalene sulfonate; tridecyl alcohol ethoxylate, POE-18;        Triethanolamine isodecanol phosphate ester; Triethanolamine        tristyrylphosphate ester; Tristyrylphenol Ethoxylate Sulfate;        Bis(2-hydroxyethyl)tallowalkylamines.    -   (e) Glucose or glucose combined with at least one material        selected from the group consisting of: lactose monohydrate;        lactose anhydrous; mannitol; microcrystalline cellulose;        sucrose; sodium chloride; talc; kaolin; calcium carbonate; malic        acid; tartaric acid; trisodium citrate dihydrate; D,L-Malic        acid; sodium pentane sulfate; sodium octadecyl sulfate; Brij700;        Brij76; sodium n-lauroyl sacrosine; lecithin; docusate sodium;        polyoxyl-40-stearate; Aerosil R972 fumed silica; sodium lauryl        sulfate or other alkyl sulfate surfactants with a chain length        between C5 to C18; polyvinyl pyrrolidone; sodium lauryl sulfate        and polyethylene glycol 40 stearate, sodium lauryl sulfate and        polyethylene glycol 100 stearate, sodium lauryl sulfate and PEG        3000, sodium lauryl sulphate and PEG 6000, sodium lauryl        sulphate and PEG 8000, sodium lauryl sulphate and PEG 10000,        sodium lauryl sulfate and Brij700, sodium lauryl sulfate and        Poloxamer 407, sodium lauryl sulfate and Poloxamer 338, sodium        lauryl sulfate and Poloxamer 188; Poloxamer 407, Poloxamer 338,        Poloxamer 188, alkyl naphthalene sulfonate        condensate/Lignosulfonate blend; Calcium Dodecylbenzene        Sulfonate (Branched); Diisopropyl naphthalenesulphonate;        erythritol distearate; linear and branched dodecylbenzene        sulfonic acids; Naphthalene Sulfonate Formaldehyde Condensate;        nonylphenol ethoxylate, POE-30; Phosphate Esters,        Tristyrylphenol Ethoxylate, Free Acid; Polyoxyethylene (15)        tallowalkylamines; sodium alkyl naphthalene sulfonate; sodium        alkyl naphthalene sulfonate condensate; sodium alkylbenzene        sulfonate; sodium isopropyl naphthalene sulfonate; Sodium Methyl        Naphthalene; Formaldehyde Sulfonate; sodium salt of n-butyl        naphthalene sulfonate; tridecyl alcohol ethoxylate, POE-18;        Triethanolamine isodecanol phosphate ester; Triethanolamine        tristyrylphosphate ester; Tristyrylphenol Ethoxylate Sulfate;        Bis(2-hydroxyethyl)tallowalkylamines.    -   (f) Sodium chloride or sodium chloride combined with at least        one material selected from the group consisting of: lactose        monohydrate; lactose anhydrous; mannitol; microcrystalline        cellulose; sucrose; glucose; talc; kaolin; calcium carbonate;        malic acid; tartaric acid; trisodium citrate dihydrate;        D,L-Malic acid; sodium pentane sulfate; sodium octadecyl        sulfate; Brij700; Brij76; sodium n-lauroyl sacrosine; lecithin;        docusate sodium; polyoxyl-40-stearate; Aerosil R972 fumed        silica; sodium lauryl sulfate or other alkyl sulfate surfactants        with a chain length between C5 to C18; polyvinyl pyrrolidone;        sodium lauryl sulfate and polyethylene glycol 40 stearate,        sodium lauryl sulfate and polyethylene glycol 100 stearate,        sodium lauryl sulfate and PEG 3000, sodium lauryl sulphate and        PEG 6000, sodium lauryl sulphate and PEG 8000, sodium lauryl        sulphate and PEG 10000, sodium lauryl sulfate and Brij700,        sodium lauryl sulfate and Poloxamer 407, sodium lauryl sulfate        and Poloxamer 338, sodium lauryl sulfate and Poloxamer 188;        Poloxamer 407, Poloxamer 338, Poloxamer 188, alkyl naphthalene        sulfonate condensate/Lignosulfonate blend; Calcium        Dodecylbenzene Sulfonate (Branched); Diisopropyl        naphthalenesulphonate; erythritol distearate; linear and        branched dodecylbenzene sulfonic acids; Naphthalene Sulfonate        Formaldehyde Condensate; nonylphenol ethoxylate, POE-30;        Phosphate Esters, Tristyrylphenol Ethoxylate, Free Acid;        Polyoxyethylene (15) tallowalkylamines; sodium alkyl naphthalene        sulfonate; sodium alkyl naphthalene sulfonate condensate; sodium        alkylbenzene sulfonate; sodium isopropyl naphthalene sulfonate;        Sodium Methyl Naphthalene; Formaldehyde Sulfonate; sodium salt        of n-butyl naphthalene sulfonate; tridecyl alcohol ethoxylate,        POE-18; Triethanolamine isodecanol phosphate ester;        Triethanolamine tristyrylphosphate ester; Tristyrylphenol        Ethoxylate Sulfate; Bis(2-hydroxyethyl)tallowalkylamines.    -   (g) xylitol or xylitol combined with at least one material        selected from the group consisting of: lactose monohydrate;        lactose anhydrous; mannitol; microcrystalline cellulose;        sucrose; glucose; sodium chloride; talc; kaolin; calcium        carbonate; malic acid; tartaric acid; trisodium citrate        dihydrate; D,L-Malic acid; sodium pentane sulfate; sodium        octadecyl sulfate; Brij700; Brij76; sodium n-lauroyl sacrosine;        lecithin; docusate sodium; polyoxyl-40-stearate; Aerosil R972        fumed silica; sodium lauryl sulfate or other alkyl sulfate        surfactants with a chain length between C5 to C18; polyvinyl        pyrrolidone; sodium lauryl sulfate and polyethylene glycol 40        stearate, sodium lauryl sulfate and polyethylene glycol 100        stearate, sodium lauryl sulfate and PEG 3000, sodium lauryl        sulphate and PEG 6000, sodium lauryl sulphate and PEG 8000,        sodium lauryl sulphate and PEG 10000, sodium lauryl sulfate and        Brij700, sodium lauryl sulfate and Poloxamer 407, sodium lauryl        sulfate and Poloxamer 338, sodium lauryl sulfate and Poloxamer        188; Poloxamer 407, Poloxamer 338, Poloxamer 188, alkyl        naphthalene sulfonate condensate/Lignosulfonate blend; Calcium        Dodecylbenzene Sulfonate (Branched); Diisopropyl        naphthalenesulphonate; erythritol distearate; linear and        branched dodecylbenzene sulfonic acids; Naphthalene Sulfonate        Formaldehyde Condensate; nonylphenol ethoxylate, POE-30;        Phosphate Esters, Tristyrylphenol Ethoxylate, Free Acid;        Polyoxyethylene (15) tallowalkylamines; sodium alkyl naphthalene        sulfonate; sodium alkyl naphthalene sulfonate condensate; sodium        alkylbenzene sulfonate; sodium isopropyl naphthalene sulfonate;        Sodium Methyl Naphthalene; Formaldehyde Sulfonate; sodium salt        of n-butyl naphthalene sulfonate; tridecyl alcohol ethoxylate,        POE-18; Triethanolamine isodecanol phosphate ester;        Triethanolamine tristyrylphosphate ester; Tristyrylphenol        Ethoxylate Sulfate; Bis(2-hydroxyethyl)tallowalkylamines.    -   (h) Tartaric acid or tartaric acid combined with at least one        material selected from the group consisting of: lactose        monohydrate; lactose anhydrous; mannitol; microcrystalline        cellulose; sucrose; glucose; sodium chloride; talc; kaolin;        calcium carbonate; malic acid; trisodium citrate dihydrate;        D,L-Malic acid; sodium pentane sulfate; sodium octadecyl        sulfate; Brij700; Brij76; sodium n-lauroyl sacrosine; lecithin;        docusate sodium; polyoxyl-40-stearate; Aerosil R972 fumed        silica; sodium lauryl sulfate or other alkyl sulfate surfactants        with a chain length between C5 to C18; polyvinyl pyrrolidone;        sodium lauryl sulfate and polyethylene glycol 40 stearate,        sodium lauryl sulfate and polyethylene glycol 100 stearate,        sodium lauryl sulfate and PEG 3000, sodium lauryl sulphate and        PEG 6000, sodium lauryl sulphate and PEG 8000, sodium lauryl        sulphate and PEG 10000, sodium lauryl sulfate and Brij700,        sodium lauryl sulfate and Poloxamer 407, sodium lauryl sulfate        and Poloxamer 338, sodium lauryl sulfate and Poloxamer 188;        Poloxamer 407, Poloxamer 338, Poloxamer 188, alkyl naphthalene        sulfonate condensate/Lignosulfonate blend; Calcium        Dodecylbenzene Sulfonate (Branched); Diisopropyl        naphthalenesulphonate; erythritol distearate; linear and        branched dodecylbenzene sulfonic acids; Naphthalene Sulfonate        Formaldehyde Condensate; nonylphenol ethoxylate, POE-30;        Phosphate Esters, Tristyrylphenol Ethoxylate, Free Acid;        Polyoxyethylene (15) tallowalkylamines; sodium alkyl naphthalene        sulfonate; sodium alkyl naphthalene sulfonate condensate; sodium        alkylbenzene sulfonate; sodium isopropyl naphthalene sulfonate;        Sodium Methyl Naphthalene; Formaldehyde Sulfonate; sodium salt        of n-butyl naphthalene sulfonate; tridecyl alcohol ethoxylate,        POE-18; Triethanolamine isodecanol phosphate ester;        Triethanolamine tristyrylphosphate ester; Tristyrylphenol        Ethoxylate Sulfate; Bis(2-hydroxyethyl)tallowalkylamines.    -   (i) microcrystalline cellulose or microcrystalline cellulose        combined with at least one material selected from the group        consisting of: lactose monohydrate; xylitol; lactose anhydrous;        mannitol; sucrose; glucose; sodium chloride; talc; kaolin;        calcium carbonate; malic acid; tartaric acid; trisodium citrate        dihydrate; D,L-Malic acid; sodium pentane sulfate; sodium        octadecyl sulfate; Brij700; Brij76; sodium n-lauroyl sacrosine;        lecithin; docusate sodium; polyoxyl-40-stearate; Aerosil R972        fumed silica; sodium lauryl sulfate or other alkyl sulfate        surfactants with a chain length between C5 to C18; polyvinyl        pyrrolidone; sodium lauryl sulfate and polyethylene glycol 40        stearate, sodium lauryl sulfate and polyethylene glycol 100        stearate, sodium lauryl sulfate and PEG 3000, sodium lauryl        sulphate and PEG 6000, sodium lauryl sulphate and PEG 8000,        sodium lauryl sulphate and PEG 10000, sodium lauryl sulfate and        Brij700, sodium lauryl sulfate and Poloxamer 407, sodium lauryl        sulfate and Poloxamer 338, sodium lauryl sulfate and Poloxamer        188; Poloxamer 407, Poloxamer 338, Poloxamer 188, alkyl        naphthalene sulfonate condensate/Lignosulfonate blend; Calcium        Dodecylbenzene Sulfonate (Branched); Diisopropyl        naphthalenesulphonate; erythritol distearate; linear and        branched dodecylbenzene sulfonic acids; Naphthalene Sulfonate        Formaldehyde Condensate; nonylphenol ethoxylate, POE-30;        Phosphate Esters, Tristyrylphenol Ethoxylate, Free Acid;        Polyoxyethylene (15) tallowalkylamines; sodium alkyl naphthalene        sulfonate; sodium alkyl naphthalene sulfonate condensate; sodium        alkylbenzene sulfonate; sodium isopropyl naphthalene sulfonate;        Sodium Methyl Naphthalene; Formaldehyde Sulfonate; sodium salt        of n-butyl naphthalene sulfonate; tridecyl alcohol ethoxylate,        POE-18; Triethanolamine isodecanol phosphate ester;        Triethanolamine tristyrylphosphate ester; Tristyrylphenol        Ethoxylate Sulfate; Bis(2-hydroxyethyl)tallowalkylamines.    -   (j) Kaolin combined with at least one material selected from the        group consisting of: lactose monohydrate; xylitol; lactose        anhydrous; mannitol; microcrystalline cellulose; sucrose;        glucose; sodium chloride; talc; kaolin; calcium carbonate; malic        acid; tartaric acid; trisodium citrate dihydrate; D,L-Malic        acid; sodium pentane sulfate; sodium octadecyl sulfate; Brij700;        Brij76; sodium n-lauroyl sacrosine; lecithin; docusate sodium;        polyoxyl-40-stearate; Aerosil R972 fumed silica; sodium lauryl        sulfate or other alkyl sulfate surfactants with a chain length        between C5 to C18; polyvinyl pyrrolidone; sodium lauryl sulfate        and polyethylene glycol 40 stearate, sodium lauryl sulfate and        polyethylene glycol 100 stearate, sodium lauryl sulfate and PEG        3000, sodium lauryl sulphate and PEG 6000, sodium lauryl        sulphate and PEG 8000, sodium lauryl sulphate and PEG 10000,        sodium lauryl sulfate and Brij700, sodium lauryl sulfate and        Poloxamer 407, sodium lauryl sulfate and Poloxamer 338, sodium        lauryl sulfate and Poloxamer 188; Poloxamer 407, Poloxamer 338,        Poloxamer 188, alkyl naphthalene sulfonate        condensate/Lignosulfonate blend; Calcium Dodecylbenzene        Sulfonate (Branched); Diisopropyl naphthalenesulphonate;        erythritol distearate; linear and branched dodecylbenzene        sulfonic acids; Naphthalene Sulfonate Formaldehyde Condensate;        nonylphenol ethoxylate, POE-30; Phosphate Esters,        Tristyrylphenol Ethoxylate, Free Acid; Polyoxyethylene (15)        tallowalkylamines; sodium alkyl naphthalene sulfonate; sodium        alkyl naphthalene sulfonate condensate; sodium alkylbenzene        sulfonate; sodium isopropyl naphthalene sulfonate; Sodium Methyl        Naphthalene; Formaldehyde Sulfonate; sodium salt of n-butyl        naphthalene sulfonate; tridecyl alcohol ethoxylate, POE-18;        Triethanolamine isodecanol phosphate ester; Triethanolamine        tristyrylphosphate ester; Tristyrylphenol Ethoxylate Sulfate;        Bis(2-hydroxyethyl)tallowalkylamines.    -   (k) Talc combined with at least one material selected from the        group consisting of: lactose monohydrate; xylitol; lactose        anhydrous; mannitol; microcrystalline cellulose; sucrose;        glucose; sodium chloride; kaolin; calcium carbonate; malic acid;        tartaric acid; trisodium citrate dihydrate; D,L-Malic acid;        sodium pentane sulfate; sodium octadecyl sulfate; Brij700;        Brij76; sodium n-lauroyl sacrosine; lecithin; docusate sodium;        polyoxyl-40-stearate; Aerosil R972 fumed silica; sodium lauryl        sulfate or other alkyl sulfate surfactants with a chain length        between C5 to C18; polyvinyl pyrrolidone; sodium lauryl sulfate        and polyethylene glycol 40 stearate, sodium lauryl sulfate and        polyethylene glycol 100 stearate, sodium lauryl sulfate and PEG        3000, sodium lauryl sulphate and PEG 6000, sodium lauryl        sulphate and PEG 8000, sodium lauryl sulphate and PEG 10000,        sodium lauryl sulfate and Brij700, sodium lauryl sulfate and        Poloxamer 407, sodium lauryl sulfate and Poloxamer 338, sodium        lauryl sulfate and Poloxamer 188; Poloxamer 407, Poloxamer 338,        Poloxamer 188, alkyl naphthalene sulfonate        condensate/Lignosulfonate blend; Calcium Dodecylbenzene        Sulfonate (Branched); Diisopropyl naphthalenesulphonate;        erythritol distearate; linear and branched dodecylbenzene        sulfonic acids; Naphthalene Sulfonate Formaldehyde Condensate;        nonylphenol ethoxylate, POE-30; Phosphate Esters,        Tristyrylphenol Ethoxylate, Free Acid; Polyoxyethylene (15)        tallowalkylamines; sodium alkyl naphthalene sulfonate; sodium        alkyl naphthalene sulfonate condensate; sodium alkylbenzene        sulfonate; sodium isopropyl naphthalene sulfonate; Sodium Methyl        Naphthalene; Formaldehyde Sulfonate; sodium salt of n-butyl        naphthalene sulfonate; tridecyl alcohol ethoxylate, POE-18;        Triethanolamine isodecanol phosphate ester; Triethanolamine        tristyrylphosphate ester; Tristyrylphenol Ethoxylate Sulfate;        Bis(2-hydroxyethyl)tallowalkylamines.

Preferably, the grinding matrix is selected from the group consistingof: a material considered to be Generally Regarded as Safe (GRAS) forpharmaceutical products; a material considered acceptable for use in anagricultural formulation; and a material considered acceptable for usein a veterinary formulation.

In another preferred embodiment, a milling aid or combination of millingaids is used. Preferably, the milling aid is selected from the groupconsisting of: colloidal silica, a surfactant, a polymer, a stearic acidand derivatives thereof. Preferably, the surfactant is in a solid formor can be manufactured into a solid form. Preferably, the surfactant isselected from the group consisting of: polyoxyethylene alkyl ethers,polyoxyethylene stearates, polyethylene glycols (PEG), poloxamers,poloxamines, sarcosine based surfactants, polysorbates, aliphaticalcohols, alkyl and aryl sulfates, alkyl and aryl polyether sulfonatesand other sulfate surfactants, trimethyl ammonium based surfactants,lecithin and other phospholipids, bile salts, polyoxyethylene castor oilderivatives, polyoxyethylene sorbitan fatty acid esters, Sorbitan fattyacid esters, Sucrose fatty acid esters, alkyl glucopyranosides, alkylmaltopyranosides, glycerol fatty acid esters, Alkyl Benzene SulphonicAcids, Alkyl Ether Carboxylic Acids, Alkyl and aryl Phosphate esters,Alkyl and aryl Sulphate esters, Alkyl and aryl Sulphonic acids, AlkylPhenol Phosphates esters, Alkyl Phenol Sulphates esters, Alkyl and ArylPhosphates, Alkyl Polysaccharides, Alkylamine Ethoxylates,Alkyl-Naphthalene Sulphonates formaldehyde condensates, Sulfosuccinates,lignosulfonates, Ceto-Oleyl Alcohol Ethoxylates, Condensed NaphthaleneSulphonates, Dialkyl and Alkyl Naphthalene Sulphonates, Di-alkylSulphosuccinates, Ethoxylated nonylphenols, Ethylene Glycol Esters,Fatty Alcohol Alkoxylates, Hydrogenated tallowalkylamines, Mono-alkylSulphosuccinamates, Nonyl Phenol Ethoxylates, Sodium Oleyl N-methylTaurate, Tallowalkylamines, linear and branched dodecylbenzene sulfonicacids.

Preferably, the surfactant is selected from the group consisting of:sodium lauryl sulfate, sodium stearyl sulfate, sodium cetyl sulfate,sodium cetostearyl sulfate, sodium docusate, sodium deoxycholate,N-lauroylsarcosine sodium salt, glyceryl monostearate, glyceroldistearate glyceryl palmitostearate, glyceryl behenate, glycerylcaprylate, glyceryl oleate, benzalkonium chloride, CTAB, CTAC,Cetrimide, cetylpyridinium chloride, cetylpyridinium bromide,benzethonium chloride, PEG 40 stearate, PEG 100 stearate, poloxamer 188,poloxamer 338, poloxamer 407 polyoxyl 2 stearyl ether, polyoxyl 100stearyl ether, polyoxyl 20 stearyl ether, polyoxyl 10 stearyl ether,polyoxyl 20 cetyl ether, polysorbate 20, polysorbate 40, polysorbate 60,polysorbate 61, polysorbate 65, polysorbate 80, polyoxyl 35 castor oil,polyoxyl 40 castor oil, polyoxyl 60 castor oil, polyoxyl 100 castor oil,polyoxyl 200 castor oil, polyoxyl 40 hydrogenated castor oil, polyoxyl60 hydrogenated castor oil, polyoxyl 100 hydrogenated castor oil,polyoxyl 200 hydrogenated castor oil, cetostearyl alcohol, macrogel 15hydroxystearate, sorbitan monopalmitate, sorbitan monostearate, sorbitantrioleate, Sucrose Palmitate, Sucrose Stearate, Sucrose Distearate,Sucrose laurate, Glycocholic acid, sodium Glycholate, Cholic Acid,Sodium Cholate, Sodium Deoxycholate, Deoxycholic acid, Sodiumtaurocholate, taurocholic acid, Sodium taurodeoxycholate,taurodeoxycholic acid, soy lecithin, phosphatidylcholine,phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol,PEG4000, PEG6000, PEG8000, PEG10000, PEG20000, alkyl naphthalenesulfonate condensate/Lignosulfonate blend, Calcium DodecylbenzeneSulfonate, Sodium Dodecylbenzene Sulfonate, Diisopropylnaphthaenesulphonate, erythritol distearate, Naphthalene SulfonateFormaldehyde Condensate, nonylphenol ethoxylate (poe-30),Tristyrylphenol Ethoxylate, Polyoxyethylene (15) tallowalkylamines,sodium alkyl naphthalene sulfonate, sodium alkyl naphthalene sulfonatecondensate, sodium alkylbenzene sulfonate, sodium isopropyl naphthalenesulfonate, Sodium Methyl Naphthalene Formaldehyde Sulfonate, sodiumn-butyl naphthalene sulfonate, tridecyl alcohol ethoxylate (poe-18),Triethanolamine isodecanol phosphate ester, Triethanolaminetristyrylphosphate ester, Tristyrylphenol Ethoxylate Sulfate,Bis(2-hydroxyethyl)tallowalkylamines. Preferably the polymer is selectedfrom the list of: polyvinylpyrrolidones (PVP), polyvinylalcohol, Acrylicacid based polymers and copolymers of acrylic acid

Preferably, the milling aid has a concentration selected from the groupconsisting of: 0.1-10% w/w, 0.1-5% w/w, 0.1-2.5% w/w, of 0.1-2% w/w,0.1-1%, 0.5-5% w/w, 0.5-3% w/w, 0.5-2% w/w, 0.5-1.5%, 0.5-1% w/w, of0.75-1.25% w/w, 0.75-1% and 1% w/w.

In another preferred embodiment of the invention, a facilitating agentis used or combination of facilitating agents is used. Preferably, thefacilitating agent is selected from the group consisting of:surfactants, polymers, binding agents, filling agents, lubricatingagents, sweeteners, flavouring agents, preservatives, buffers, wettingagents, disintegrants, effervescent agents, agents that may form part ofa medicament, including a solid dosage form or a dry powder inhalationformulation and other material required for specific drug delivery.Preferably, the facilitating agent is added during dry milling.Preferably, the facilitating agent is added to the dry milling at a timeselected from the group consisting of: with 1-5% of the total millingtime remaining, with 1-10% of the total milling time remaining, with1-20% of the total milling time remaining, with 1-30% of the totalmilling time remaining, with 2-5% of the total milling time remaining,with 2-10% of the total milling time remaining, with 5-20% of the totalmilling time remaining and with 5-20% of the total milling timeremaining. Preferably, the disintegrant is selected from the groupconsisting of: crosslinked PVP, cross linked carmellose and sodiumstarch glycolate. Preferably, the facilitating agent is added to themilled biologically active material and grinding matrix and furtherprocessed in a mechanofusion process. Mechanofusion milling causesmechanical energy to be applied to powders or mixtures of particles inthe micrometre and nanometre range.

The reasons for including facilitating agents include, but are notlimited to providing better dispersibility, control of agglomeration,the release or retention of the active particles from the deliverymatrix. Examples of facilitating agents include, but are not limited tocrosslinked PVP (crospovidone), cross linked carmellose(croscarmellose), sodium starch glycolate, Povidone (PVP), Povidone K12,Povidone K17, Povidone K25, Povidone K29/32 and Povidone K30, stearicacid, magnesium stearate, calcium stearate, sodium stearyl fumarate,sodium stearyl lactylate, zinc stearate, sodium stearate or lithiumstearate, other solid state fatty acids such as oleic acid, lauric acid,palmitic acid, erucic acid, behenic acid, or derivatives (such as estersand salts), Amino acids such as leucine, isoleucine, lysine, valine,methionine, phenylalanine, aspartame or acesulfame K. In a preferredaspect of manufacturing this formulation the facilitating agent is addedto the milled mixture of biologically active material and co-grindingmatrix and further processed in another milling device such asMechnofusion, Cyclomixing, or impact milling such as ball milling, jetmilling, or milling using a high pressure homogeniser, or combinationsthereof. In a highly preferred aspect the facilitating agent is added tothe milling of the mixture of biologically active material andco-grinding matrix as some time before the end of the milling process.

In another preferred embodiment, naproxen is milled with lactosemonohydrate and alkyl sulfates. Preferably naproxen is milled withlactose monohydrate and sodium lauryl sulfate. Preferably naproxen ismilled with lactose monohydrate and sodium octadecyl sulfate. In anotherpreferred embodiment, Naproxen is milled with lactose monohydrate, alkylsulfates and another surfactant or polymers. Preferably naproxen ismilled with lactose monohydrate, sodium lauryl sulfate and polyethersulfates. Preferably naproxen is milled with lactose monohydrate, sodiumlauryl sulfate and polyethylene glycol 40 stearate. Preferably naproxenis milled with lactose monohydrate, sodium lauryl sulfate andpolyethylene glycol 100 stearate. Preferably naproxen is milled withlactose monohydrate, sodium lauryl sulfate and a poloxamer. Preferablynaproxen is milled with lactose monohydrate, sodium lauryl sulfate andpoloxamer 407. Preferably naproxen is milled with lactose monohydrate,sodium lauryl sulfate and poloxamer 338. Preferably naproxen is milledwith lactose monohydrate, sodium lauryl sulfate and poloxamer 188.Preferably naproxen is milled with lactose monohydrate, sodium laurylsulfate and a solid polyethylene glycol. Preferably naproxen is milledwith lactose monohydrate, sodium lauryl sulfate and polyethylene glycol6000. Preferably naproxen is milled with lactose monohydrate, sodiumlauryl sulfate and polyethylene glycol 3000. In another preferredembodiment, Naproxen is milled with lactose monohydrate and polyethersulfates. Preferably naproxen is milled with lactose monohydrate andpolyethylene glycol 40 stearate. Preferably naproxen is milled withlactose monohydrate and polyethylene glycol 100 stearate In anotherpreferred embodiment naproxen is milled with lactose monohydrate andpolyvinyl-pyrrolidine. Preferably naproxen is milled with lactosemonohydrate and polyvinyl-pyrrolidone with an approximate molecularweight of 30,000-40,000. In another preferred embodiment, naproxen ismilled with lactose monohydrate and alkyl sulfonates. Preferablynaproxen is milled with lactose monohydrate and docusate sodium. Inanother preferred embodiment, naproxen is milled with lactosemonohydrate and a surfactant. Preferably naproxen is milled with lactosemonohydrate and lecithin. Preferably naproxen is milled with lactosemonohydrate and sodium n-lauroyl sarcosine. Preferably naproxen ismilled with lactose monohydrate and polyoxyethylene alkyl ethersurfactants. Preferably naproxen is milled with lactose monohydrate andPEG 6000. In another preferred formulation naproxen is milled withlactose monohydrate and silica. Preferably naproxen is milled withlactose monohydrate and Aerosil R972 fumed silica. In another preferredembodiment, naproxen is milled with lactose monohydrate, tartaric acidand sodium lauryl sulfate. In another preferred embodiment, naproxen ismilled with lactose monohydrate, sodium bicarbonate and sodium laurylsulfate. In another preferred embodiment, naproxen is milled withlactose monohydrate, sodium bicarbonate, poloxamer 407 and sodium laurylsulfate. In another preferred embodiment, naproxen is milled withlactose monohydrate, potassium bicarbonate and sodium lauryl sulfate. Inanother preferred embodiment, naproxen is milled with lactosemonohydrate, potassium bicarbonate, poloxamer 407 and sodium laurylsulfate. In another preferred embodiment, naproxen is milled withmannitol and alkyl sulfates. Preferably naproxen is milled with mannitoland sodium lauryl sulfate. Preferably naproxen is milled with mannitoland sodium octadecyl sulfate. In another preferred embodiment, Naproxenis milled with mannitol, alkyl sulfates and another surfactant orpolymers. Preferably naproxen is milled with mannitol, sodium laurylsulfate and polyether sulfates. Preferably naproxen is milled withmannitol, sodium lauryl sulfate and polyethylene glycol 40 stearate.Preferably naproxen is milled with mannitol, sodium lauryl sulfate andpolyethylene glycol 100 stearate. Preferably naproxen is milled withmannitol, sodium lauryl sulfate and a poloxamer. Preferably naproxen ismilled with mannitol, sodium lauryl sulfate and poloxamer 407.Preferably naproxen is milled with mannitol, sodium lauryl sulfate andpoloxamer 338. Preferably naproxen is milled with mannitol, sodiumlauryl sulfate and poloxamer 188. Preferably naproxen is milled withmannitol, sodium lauryl sulfate and a solid polyethylene glycol.Preferably naproxen is milled with mannitol, sodium lauryl sulfate andpolyethylene glycol 6000. Preferably naproxen is milled with mannitol,sodium lauryl sulfate and polyethylene glycol 3000. In another preferredembodiment, Naproxen is milled with mannitol and polyether sulfates.Preferably naproxen is milled with mannitol and polyethylene glycol 40stearate. Preferably naproxen is milled with mannitol and polyethyleneglycol 100 stearate In another preferred embodiment naproxen is milledwith mannitol and polyvinyl-pyrrolidine. Preferably naproxen is milledwith mannitol and polyvinyl-pyrrolidone with an approximate molecularweight of 30,000-40,000. In another preferred embodiment, naproxen ismilled with mannitol and alkyl sulfonates. Preferably naproxen is milledwith mannitol and docusate sodium. In another preferred embodiment,naproxen is milled with mannitol and a surfactant. Preferably naproxenis milled with mannitol and lecithin. Preferably naproxen is milled withmannitol and sodium n-lauroyl sarcosine. Preferably naproxen is milledwith mannitol and polyoxyethylene alkyl ether surfactants. Preferablynaproxen is milled with mannitol and PEG 6000. In another preferredformulation naproxen is milled with mannitol and silica. Preferablynaproxen is milled with mannitol and Aerosil R972 fumed silica. Inanother preferred embodiment, naproxen is milled with mannitol, tartaricacid and sodium lauryl sulfate. In another preferred embodiment,naproxen is milled with mannitol, sodium bicarbonate and sodium laurylsulfate. In another preferred embodiment, naproxen is milled withmannitol, potassium bicarbonate and sodium lauryl sulfate. In anotherpreferred embodiment, naproxen is milled with mannitol, sodiumbicarbonate and sodium lauryl sulphate and Polxamer 407. In anotherpreferred embodiment, naproxen is milled with mannitol, potassiumbicarbonate and sodium lauryl sulphate and Polxamer 407.

In a second aspect the invention comprises a biologically activematerial produced by the method described herein and compositioncomprising the biologically active material as described herein.Preferably, the average particle size, determined on a particle numberbasis, is equal to or less than a size selected from the group 2000 nm,1900 nm, 1800 nm, 1700 nm, 1600 nm, 1500 nm, 1400 nm, 1300 nm, 1200 nm,1100 nm, 1000 nm, 900 nm, 800 nm, 700 nm, 600 nm, 500 nm, 400 nm, 300nm, 200 nm and 100 nm. Preferably, the average particle size is equal toor greater than 25 nm. Preferably, the particles have a median particlesize, determined on a particle volume basis, equal or less than a sizeselected from the group 2000 nm, 1900 nm, 1800 nm, 1700 nm, 1600 nm,1500 nm, 1400 nm, 1300 nm, 1200 nm, 1100 nm, 1000 nm, 900 nm, 800 nm,700 nm, 600 nm, 500 nm, 400 nm, 300 nm, 200 nm and 100 nm. Preferably,the median particle size is equal to or greater than 25 nm. Preferably,the percentage of particles, on a particle volume basis, is selectedfrom the group consisting of: 50%, 60%, 70%, 80%, 90%, 95% and 100% lessthan 2000 nm (%<2000 nm). Preferably, the percentage of particles, on aparticle volume basis, is selected from the group consisting of: 50%,60%, 70%, 80%, 90%, 95% and 100% less than 1000 nm (%<1000 nm).Preferably, the percentage of particles, on a particle volume basis, isselected from the group 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%,95% and 100% less than 500 nm (%<500 nm). Preferably, the percentage ofparticles, on a particle volume basis, is selected from the group 0%,10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% and 100% less than 300nm (%<300 nm). Preferably, the percentage of particles, on a particlevolume basis, is selected from the group 0%, 10%, 20%, 30%, 40%, 50%,60%, 70%, 80%, 90%, 95% and 100% less than 200 nm (%<200 nm).Preferably, the Dx of the particle size distribution, as measured on aparticle volume basis, is selected from the group consisting of lessthan or equal to 10,000 nm, 5000 nm, 3000 nm, 2000 nm, 1900 nm, 1800 nm,1700 nm, 1600 nm, 1500 nm, 1400 nm, 1300 nm, 1200 nm, 1100 nm, 1000 nm,900 nm, 800 nm, 700 nm, 600 nm, 500 nm, 400 nm, 300 nm, 200 nm, and 100nm; wherein x is greater than or equal to 90. Preferably, thebiologically active material comprised in the composition is naproxen orany salt or derivative thereof.

In one preferred embodiment, the invention comprises compositionscomprising the biologically active ingredient together with a grindingmatrix, a mixture of grinding matrix materials, milling aids, mixturesof milling aids, facilitating agents and/or mixtures of facilitatingagents as described herein, in concentrations and ratios as describedherein under the methods of the invention.

In a third aspect the invention comprises a pharmaceutical compositioncomprising a biologically active material produced by the methoddescribed herein and compositions described herein. Preferably, theinvention comprises pharmaceutical compositions comprising thebiologically active ingredient together with a grinding matrix, amixture of grinding matrix materials, milling aids, mixtures of millingaids, facilitating agents and/or mixtures of facilitating agents asdescribed herein, in concentrations and ratios as described herein underthe methods of the invention. Preferably, the average particle size,determined on a particle number basis, is equal to or less than a sizeselected from the group 2000 nm, 1900 nm, 1800 nm, 1700 nm, 1600 nm,1500 nm, 1400 nm, 1300 nm, 1200 nm, 1100 nm, 1000 nm, 900 nm, 800 nm,700 nm, 600 nm, 500 nm, 400 nm, 300 nm, 200 nm and 100 nm. Preferably,the average particle size is equal to or greater than 25 nm. Preferably,the particles have a median particle size, determined on a particlevolume basis, equal or less than a size selected from the group 2000 nm,1900 nm, 1800 nm, 1700 nm, 1600 nm, 1500 nm, 1400 nm, 1300 nm, 1200 nm,1100 nm, 1000 nm, 900 nm, 800 nm, 700 nm, 600 nm, 500 nm, 400 nm, 300nm, 200 nm and 100 nm. Preferably, the median particle size is equal toor greater than 25 nm. Preferably, the percentage of particles, on aparticle volume basis, is selected from the group consisting of: lessthan 2000 nm (%<2000 nm) is selected from the group consisting of: 50%,60%, 70%, 80%, 90%, 95% and 100%; less than 1000 nm (%<1000 nm) isselected from the group consisting of: 50%, 60%, 70%, 80%, 90%, 95% and100%; less than 500 nm (%<500 nm) is selected from the group 0%, 10%,20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% and 100%; less than 300 nm(%<300 nm) is selected from the group 0%, 10%, 20%, 30%, 40%, 50%, 60%,70%, 80%, 90%, 95% and 100%; and less than 200 nm (%<200 nm) is selectedfrom the group 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% and100%.

Preferably, the crystallinity profile of the biologically activematerial is selected from the group consisting of: at least 50% of thebiologically active material is crystalline, at least 60% of thebiologically active material is crystalline, at least 70% of thebiologically active material is crystalline, at least 75% of thebiologically active material is crystalline, at least 85% of thebiologically active material is crystalline, at least 90% of thebiologically active material is crystalline, at least 95% of thebiologically active material is crystalline and at least 98% of thebiologically active material is crystalline. Preferably, thecrystallinity profile of the biologically active material issubstantially equal to the crystallinity profile of the biologicallyactive material before the material was subject to the method describedherein. Preferably, the amorphous content of the biologically activematerial is selected from the group consisting of: less than 50% of thebiologically active material is amorphous, less than 40% of thebiologically active material is amorphous, less than 30% of thebiologically active material is amorphous, less than 25% of thebiologically active material is amorphous, less than 15% of thebiologically active material is amorphous, less than 10% of thebiologically active material is amorphous, less than 5% of thebiologically active material is amorphous and less than 2% of thebiologically active material is amorphous. Preferably, the biologicallyactive material has had no significant increase in amorphous contentfollowing subjecting the material to the method as described herein.

Preferably, the biologically active material is naproxen or derivativesor salts thereof. Preferably, the composition has a T_(max) less thanthat of the equivalent conventional composition administered at the samedosage, wherein the composition comprises naproxen. Preferably, thecomposition has a C_(max) greater than that of the equivalentconventional composition administered at the same dosage, wherein thecomposition comprises naproxen. Preferably, the composition has an AUCgreater than that of the equivalent conventional compositionadministered at the same dosage, wherein the composition comprisesnaproxen. In a fourth aspect the invention comprises a method oftreating a human in need of such treatment comprising the step ofadministering to the human an effective amount of a pharmaceuticalcomposition as described herein.

In a fifth aspect, the invention comprises the use of a pharmaceuticalcomposition as described herein in the manufacture of a medicament forthe treatment of a human in need of such treatment.

In a sixth aspect the invention comprises a method for manufacturing apharmaceutical composition as described herein comprising the step ofcombining a therapeutically effective amount of a biologically activematerial prepared by a method described herein or a composition asdescribed herein, together with a pharmaceutically acceptable carrier toproduce a pharmaceutically acceptable dosage form.

In a seventh aspect the invention comprises a method for manufacturing aveterinary product comprising the step of combining a therapeuticallyeffective amount of the biologically active material prepared by amethod as described herein or a composition as described herein,together with an acceptable excipient to produce a dosage formacceptable for veterinary use. In an eighth aspect the inventioncomprises a method for manufacturing of a pharmaceutical formulationcomprising the step of combining an effective amount of the biologicallyactive material prepared by a method described herein together withacceptable excipients to produce a formulation that can deliver atherapeutically effective amount of active to the pulmonary or nasalarea. Such a formulation could be, but is not limited to a dry powderformulation for oral inhalation to the lungs or a formulation for nasalinhalation. Preferably the method for manufacturing such a formulationuses lactose, mannitol, sucrose, sorbitol, xylitol or other sugars orpolyols as the co-grinding matrix together with surfactant such as, butnot limited to lecithin, DPPC (dipalmitoyl phosphatidylcholine), PG(phosphatidylglycerol), dipalmitoyl phosphatidyl ethanolamine (DPPE),dipalmitoyl phosphatidylinositol (DPPI) or other phospholipid. Theparticle size of the material produced by the invention disclosed hereinresults in the materials being readily aerosolized and suitable formethods of delivery to a subject in need thereof, including pulmonaryand nasal delivery methods.

While the method of the present invention has particular application inthe preparation of poorly water-soluble biologically active materials,the scope of the invention is not limited thereto. For example, themethod of the present invention enables production of highlywater-soluble biologically active materials. Such materials may exhibitadvantages over conventional materials by way of, for example, morerapid therapeutic action or lower dose. In contrast, wet grindingtechniques utilizing water (or other comparably polar solvents) areincapable of being applied to such materials, as the particles dissolveappreciably in the solvent.

Other aspects and advantages of the invention will become apparent tothose skilled in the art from a review of the ensuing description.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A. Powder charge composition and particle size distribution ofmaterial milled in SPEX mill, examples A to S.

FIG. 1B. Powder charge composition and particle size distribution ofmaterial milled in SPEX mill, examples T to AL.

FIG. 1C. Powder charge composition and particle size distribution ofmaterial milled in SPEX mill, examples AM to BE.

FIG. 1D. Powder charge composition and particle size distribution ofmaterial milled in SPEX mill, examples BF to BX.

FIG. 1E. Powder charge composition and particle size distribution ofmaterial milled in SPEX mill, examples BY to CQ.

FIG. 1F. Powder charge composition and particle size distribution ofmaterial milled in SPEX mill, examples CR to DJ.

FIG. 1G. Powder charge composition and particle size distribution ofmaterial milled in SPEX mill, examples DK to EC.

FIG. 1H. The figure shows the X-Ray diffraction patterns: (A) aftermilling of Naproxen sodium in tartaric acid; (B) unmilled Naproxensodium and (C) unmilled Naproxen acid.

FIG. 2A. Powder charge composition and particle size distribution ofmaterial milled in 110 mL HD01 Attritor mill, examples A to F.

FIG. 3A. Powder charge composition and particle size distribution ofmaterial containing a mixture of 2 matrices, milled in SPEX mill,examples A to E.

FIG. 4A. Powder charge composition and particle size distribution ofmaterial milled in 1 L HD01 Attritor mill, examples A to G.

FIG. 5A. Powder charge composition and particle size distribution ofmaterial milled in 750 mL 1S Attritor mill, examples A to F.

FIG. 6A. Powder charge composition and particle size distribution ofmaterial milled in ½ Gallon 1S Attritor mill, examples A to R.

FIG. 6B. Powder charge composition and particle size distribution ofmaterial milled in ½ Gallon 1S Attritor mill, examples S to AK.

FIG. 6C. Powder charge composition and particle size distribution ofmaterial milled in ½ Gallon 1S Attritor mill, examples AL to AU.

FIG. 7A. Powder charge composition and particle size distribution ofNaproxen milled in a variety of mills, examples A to O.

FIG. 8A. Powder charge composition and particle size distribution ofmaterial milled in HICOM mill, examples A to P.

FIG. 9A. Powder charge composition and particle size distribution ofmaterial milled in 1½ Gallon 1S Attritor mill, examples A to S.

FIG. 9B. Powder charge composition and particle size distribution ofmaterial milled in 1½ Gallon 1S Attritor mill, examples T to AL.

FIG. 10A. Powder charge composition and particle size distribution ofmaterial milled in a variety of large scale mills, examples A to F.

FIG. 11A. Powder charge composition and particle size distribution ofNaproxen Acid milled in Mannitol in a ½ Gallon 1S Attritor mill,examples A to M.

FIG. 12A. Powder charge composition and particle size distribution ofNaproxen Acid milled in SPEX mill and particle size distribution afterfiltration, examples A to L.

DETAILED DESCRIPTION OF THE INVENTION

General

Those skilled in the art will appreciate that the invention describedherein is susceptible to variations and modifications other than thosespecifically described. It is to be understood that the inventionincludes all such variations and modifications. The invention alsoincludes all of the steps, features, compositions and materials referredto or indicated in the specification, individually or collectively andany and all combinations or any two or more of the steps or features.

The present invention is not to be limited in scope by the specificembodiments described herein, which are intended for the purpose ofexemplification only. Functionally equivalent products, compositions andmethods are clearly within the scope of the invention as describedherein.

The invention described herein may include one or more ranges of values(e.g. size, concentration etc). A range of values will be understood toinclude all values within the range, including the values defining therange, and values adjacent to the range that lead to the same orsubstantially the same outcome as the values immediately adjacent tothat value which defines the boundary to the range.

The entire disclosures of all publications (including patents, patentapplications, journal articles, laboratory manuals, books, or otherdocuments) cited herein are hereby incorporated by reference. Inclusiondoes not constitute an admission is made that any of the referencesconstitute prior art or are part of the common general knowledge ofthose working in the field to which this invention relates.

Throughout this specification, unless the context requires otherwise,the word “comprise” or variations, such as “comprises” or “comprising”will be understood to imply the inclusion of a stated integer, or groupof integers, but not the exclusion of any other integers or group ofintegers. It is also noted that in this disclosure, and particularly inthe claims and/or paragraphs, terms such as “comprises”, “comprised”,“comprising” and the like can have the meaning attributed to it in USPatent law; e.g., they can mean “includes”, “included”, “including”, andthe like.

“Therapeutically effective amount” as used herein with respect tomethods of treatment and in particular drug dosage, shall mean thatdosage that provides the specific pharmacological response for which thedrug is administered in a significant number of subjects in need of suchtreatment. It is emphasized that “therapeutically effective amount,”administered to a particular subject in a particular instance will notalways be effective in treating the diseases described herein, eventhough such dosage is deemed a “therapeutically effective amount” bythose skilled in the art. It is to be further understood that drugdosages are, in particular instances, measured as oral dosages, or withreference to drug levels as measured in blood.

The term “inhibit” is defined to include its generally accepted meaningwhich includes prohibiting, preventing, restraining, and lowering,stopping, or reversing progression or severity, and such action on aresultant symptom. As such the present invention includes both medicaltherapeutic and prophylactic administration, as appropriate.

The term “biologically active material” is defined to mean abiologically active compound or a substance which comprises abiologically active compound. In this definition, a compound isgenerally taken to mean a distinct chemical entity where a chemicalformula or formulas can be used to describe the substance. Suchcompounds would generally, but not necessarily be identified in theliterature by a unique classification system such as a CAS number. Somecompounds may be more complex and have a mixed chemical structure. Forsuch compounds they may only have an empirical formula or bequalitatively identified. A compound would generally be a pure material,although it would be expected that up to 10%, 20%, 30%, 40%, 50%, 60%,70%, 80%, 90% of the substance could be other impurities and the like.Examples of biologically active compounds are, but not limited to,pharmaceutical actives, homologs and first order derivatives thereof. Asubstance that contains a biologically active compound is any substancewhich has as one of its components a biologically active compound.Examples of substances containing biologically active compounds are, butnot limited to, pharmaceutical formulations and products.

Any of the terms, “biological(ly) active”, “active”, “active material”shall have the same meaning as biologically active material.

The term “grinding matrix” is defined as any inert substance that abiologically active material can or is combined with and milled. Theterms “co-grinding matrix” and “matrix” are interchangeable with“grinding matrix”.

Particle Size

There are a wide range of techniques that can be utilized tocharacterize the particle size of a material. Those skilled in the artalso understand that almost all these techniques do not physicallymeasure the actually particle size, as one might measure something witha ruler, but measure a physical phenomena which is interpreted toindicate a particle size. As part of the interpretation process someassumptions need to be made to enable mathematical calculations to bemade. These assumptions deliver results such as an equivalent sphericalparticle size, or a hydrodynamic radius.

Amongst these various methods, two types of measurements are mostcommonly used. Photon correlation spectroscopy (PCS), also known as‘dynamic light scattering’ (DLS) is commonly used to measure particleswith a size less than 10 micron. Typically this measurement yields anequivalent hydrodynamic radius often expressed as the average size of anumber distribution. The other common particle size measurement is laserdiffraction which is commonly used to measure particle size from 100 nmto 2000 micron. This technique calculates a volume distribution ofequivalent spherical particles that can be expressed using descriptorssuch as the median particle size or the % of particles under a givensize.

Those skilled in the art recognize that different characterizationtechniques such as photon correlation spectroscopy and laser diffractionmeasure different properties of a particle ensemble. As a resultmultiple techniques will give multiple answers to the question, “what isthe particle size.” In theory one could convert and compare the variousparameters each technique measures, however, for real world particlesystems this is not practical. As a result the particle size used todescribe this invention will be given as two different sets of valuesthat each relate to these two common measurement techniques, such thatmeasurements could be made with either technique and then evaluatedagainst the description of this invention.

For measurements made using a photo correlation spectroscopy instrument,or an equivalent method known in the art, the term “number averageparticle size” is defined as the average particle diameter as determinedon a number basis.

For measurements made using a laser diffraction instrument, or anequivalent method known in the art, the term “median particle size” isdefined as the median particle diameter as determined on an equivalentspherical particle volume basis. Where the term median is used, it isunderstood to describe the particle size that divides the population inhalf such that 50% of the population is greater than or less than thissize. The median particle size is often written as D50, D(0.50) orD[0.5] or similar. As used herein D50, D(0.50) or D[0.5] or similarshall be taken to mean ‘median particle size’.

The term “Dx of the particle size distribution” refers to the xthpercentile of the distribution; thus, D90 refers to the 90^(th)percentile, D95 refers to the 95^(th) percentile, and so forth. TakingD90 as an example this can often be written as, D(0.90) or D[0.9] orsimilar. With respect to the median particle size and Dx an upper case Dor lowercase d are interchangeable and have the same meaning.

Another commonly used way of describing a particle size distributionmeasured by laser diffraction, or an equivalent method known in the art,is to describe what % of a distribution is under or over a nominatedsize. The term “percentage less than” also written as “%<” is defined asthe percentage, by volume, of a particle size distribution under anominated size—for example the %<1000 nm. The term “percentage greaterthan” also written as “%>” is defined as the percentage, by volume, of aparticle size distribution over a nominated size—for example the %>1000nm.

The particle size used to describe this invention should be taken tomean the particle size as measured at or shortly before the time of use.For example, the particle size is measured 2 months after the materialis subject to the milling method of this invention. In a preferred form,the particle size is measured at a time selected from the groupconsisting of: 1 day after milling, 2 days after milling, 5 days aftermilling, 1 month after milling, 2 months after milling, 3 months aftermilling, 4 months after milling, 5 months after milling, 6 months aftermilling, 1 year after milling, 2 years after milling, 5 years aftermilling.

For many of the materials subject to the methods of this invention theparticle size can be easily measured. Where the active material has poorwater solubility and the matrix it is milled in has good watersolubility the powder can simply be dispersed in an aqueous solvent. Inthis scenario the matrix dissolves leaving the active material dispersedin the solvent. This suspension can then be measured by techniques suchas PCS or laser diffraction.

Suitable methods to measure an accurate particle size where the activematerial has substantive aqueous solubility or the matrix has lowsolubility in a water based dispersant are outlined below.

-   -   1. In the circumstance where insoluble matrix such as        microcrystalline cellulose prevents the measurement of the        active material separation techniques such as filtration or        centrifugation could be used to separate the insoluble matrix        from the active material particles. Other ancillary techniques        would also be required to determine if any active material was        removed by the separation technique so that this could be taken        into account.    -   2. In the case where the active material is too soluble in water        other solvents could be evaluated for the measurement of        particle size. Where a solvent could be found that active        material is poorly soluble in but is a good solvent for the        matrix a measurement would be relatively straight forward. If        such a solvent is difficult to find another approach would be to        measure the ensemble of matrix and active material in a solvent        (such as iso-octane) which both are insoluble in. Then the        powder would be measured in another solvent where the active        material is soluble but the matrix is not. Thus with a        measurement of the matrix particle size and a measurement of the        size of the matrix and active material together an understanding        of the active material particle size can be obtained.    -   3. In some circumstances image analysis could be used to obtain        information about the particle size distribution of the active        material. Suitable image measurement techniques might include        transmission electron microscopy (TEM), scanning electron        microscopy (SEM), optical microscopy and confocal microscopy. In        addition to these standard techniques some additional technique        would be required to be used in parallel to differentiate the        active material and matrix particles. Depending on the chemical        makeup of the materials involved possible techniques could be        elemental analysis, raman spectroscopy, FTIR spectroscopy or        fluorescence spectroscopy.

Other Definitions

Throughout this specification, unless the context requires otherwise,the phrase “dry mill” or variations, such as “dry milling”, should beunderstood to refer to milling in at least the substantial absence ofliquids. If liquids are present, they are present in such amounts thatthe contents of the mill retain the characteristics of a dry powder.

“Flowable” means a powder having physical characteristics rendering itsuitable for further processing using typical equipment used for themanufacture of pharmaceutical compositions and formulations.

Other definitions for selected terms used herein may be found within thedetailed description of the invention and apply throughout. Unlessotherwise defined, all other scientific and technical terms used hereinhave the same meaning as commonly understood to one of ordinary skill inthe art to which the invention belongs.

The term “millable” means that the grinding matrix is capable of beingphysically degraded under the dry milling conditions of the method ofthe invention. In one embodiment of the invention, the milled grindingmatrix is of a comparable particle size to the biologically activematerial. In another embodiment of the invention the particle size ofthe matrix is substantially reduced but not as small as the biologicallyactive material

Other definitions for selected terms used herein may be found within thedetailed description of the invention and apply throughout. Unlessotherwise defined, all other scientific and technical terms used hereinhave the same meaning as commonly understood to one of ordinary skill inthe art to which the invention belongs.

Specific

In one embodiment, the present invention is directed to a method forproducing a composition, comprising the steps of: dry milling a solidbiologically active material and a millable grinding matrix in a millcomprising a plurality of milling bodies, for a time period sufficientto produce particles of the biologically active material dispersed in anat least partially milled grinding material, wherein the compositionproduced by said method comprises particles of the biologically activecompound at or above a volume fraction of 25 v/v %.

The mixture of active material and matrix may then be separated from themilling bodies and removed from the mill.

In one aspect the mixture of active material and matrix is then furtherprocessed. In another aspect, the grinding matrix is separated from theparticles of biologically active material. In a further aspect, at leasta portion of the milled grinding matrix is separated from theparticulate biologically active material.

The milling bodies are essentially resistant to fracture and erosion inthe dry milling process. The quantity of the grinding matrix relative tothe quantity of biologically active material in particulate form, andthe extent of milling of the grinding matrix, is sufficient to inhibitre-agglomeration of the particles of the active material.

The present invention also relates to biologically active materialsproduced by said methods, to medicaments produced using saidbiologically active materials and to methods of treatment of an animal,including man, using a therapeutically effective amount of saidbiologically active materials administered by way of said medicaments.

Increasing the Volume Fraction Load

The present invention is directed to the unexpected finding thatparticles of a biologically active material can be produced by drymilling processes wherein the composition produced by said methodcomprises particles of the biologically active compound at or above avolume fraction of 25 v/v %. In one surprising aspect the particle sizeproduced by the process is equal to or less than 2000 nm. In anothersurprising aspect the particle size produced by the process is equal toor less than 1000 nm. This can result in a more efficient and costeffective process.

Improving the Dissolution Profile

The process results in the biologically active material having animproved dissolution profile. An improved dissolution profile hassignificant advantages including the improvement of bioavailability ofthe biologically active material in vivo. Preferably, the improveddissolution profile is observed in vitro. Alternatively, the improveddissolution profile is observed in vivo by the observation of animproved bioavailability profile. Standard methods for determining thedissolution profile of a material in vitro are available in the art. Asuitable method to determine an improved dissolution profile in vitromay include determining the concentration of the sample material in asolution over a period of time and comparing the results from the samplematerial to a control sample. An observation that peak solutionconcentration for the sample material was achieved in less time than thecontrol sample would indicate (assuming it is statisticallysignificant), that the sample material has an improved dissolutionprofile. The measurement sample is herein defined as the mixture ofbiologically active material with grinding matrix and/or other additivesthat has been subject to the processes of the invention described here.Herein a control sample is defined as a physical mixture (not subject tothe processes described in this invention) of the components in themeasurement sample with the same relative proportions of active, matrixand/or additive as the measurement sample. For the purposes of thedissolution testing a prototype formulation of the measurement samplecould also be used. In this case the control sample would be formulatedin the same way. Standard methods for determining the improveddissolution profile of a material in vivo are available in the art. Asuitable method to determine an improved dissolution profile in a humanmay be after delivering the dose to measure the rate of active materialabsorption by measuring the plasma concentration of the sample compoundover a period of time and comparing the results from the sample compoundto a control. An observation that peak plasma concentration for thesample compound was achieved in less time than the control wouldindicate (assuming it is statistically significant) that the samplecompound has improved bioavailability and an improved dissolutionprofile. Preferably, the improved dissolution profile is observed at arelevant gastrointestinal pH, when it is observed in vitro. Preferably,the improved dissolution profile is observed at a pH which is favourableat indicating improvements in dissolution when comparing the measurementsample to the control compound. Suitable methods for quantifying theconcentration of a compound in an in vitro sample or an in vivo sampleare widely available in the art. Suitable methods could include the useof spectroscopy or radioisotope labeling. In one preferred embodimentthe method of quantification of dissolution is determined in a solutionwith a pH selected from the group consisting of: pH 1, pH 2, pH 3, pH 4,pH 5, pH 6, pH 7, pH 7.3, pH 7.4, pH 8, pH 9, pH 10, pH 11, pH 12, pH13, pH 14 or a pH with 0.5 of a pH unit of any of this group.

Crystallization Profile

Methods for determining the crystallinity profile of the biologicallyactive material are widely available in the art. Suitable methods mayinclude X-ray diffraction, differential scanning calorimetry, raman orIR spectrocopy.

Amorphicity Profile

Methods for determining the amorphous content of the biologically activematerial are widely available in the art. Suitable methods may includeX-ray diffraction, differential scanning calorimetry, raman or IRspectroscopy.

Grinding Matrix

As will be described subsequently, selection of an appropriate grindingmatrix affords particular advantageous applications of the method of thepresent invention.

A highly advantageous application of the method of the invention is theuse of a water-soluble grinding matrix in conjunction with a poorlywater-soluble biologically active material. This affords at least twoadvantages. The first being when the powder containing the biologicallyactive material is placed into water—such as the ingestion of the powderas part of an oral medication—the matrix dissolves, releasing theparticulate active material such that there is maximum surface areaexposed to solution, thereby allowing a rapid dissolution of the activecompound. The second key advantage is the ability, if required, toremove or partially remove the matrix prior to further processing orformulation.

Another advantageous application of the method of the invention is theuse of a water-insoluble grinding matrix, particularly in the area ofagricultural use, when a biologically active material such as afungicide is commonly delivered as part of a dry powder or a suspension.The presence of a water insoluble matrix will afford benefits such asincreased rain fastness.

Without wishing to be bound by theory, it is believed that the physicaldegradation (including but not limited to particle size reduction) ofthe millable grinding matrix affords the advantage of the invention, byacting as a more effective diluent than grinding matrix of a largerparticle size.

Again, as will be described subsequently, a highly advantageous aspectof the present invention is that certain grinding matrixes appropriatefor use in the method of the invention are also appropriate for use in amedicament. The present invention encompasses methods for the productionof a medicament incorporating both the biologically active material andthe grinding matrix or in some cases the biologically active materialand a portion of the grinding matrix, medicaments so produced, andmethods of treatment of an animal, including man, using atherapeutically effective amount of said biologically active materialsby way of said medicaments.

The medicament may include only the biologically active materialtogether with the milled grinding matrix or, more preferably, thebiologically active material and milled grinding matrix may be combinedwith one or more pharmaceutically acceptable carriers, as well as anydesired excipients or other like agents commonly used in the preparationof medicaments. Analogously, the agricultural chemical composition mayinclude only the biologically active material together with the milledgrinding matrix or, more preferably, the biologically active materialsand milled grinding matrix may be combined with one or more carriers, aswell as any desired excipients or other like agents commonly used in thepreparation of agricultural chemical compositions.

In one particular form of the invention, the grinding matrix is bothappropriate for use in a medicament and readily separable from thebiologically active material by methods not dependent on particle size.Such grinding matrixes are described in the following detaileddescription of the invention. Such grinding matrixes are highlyadvantageous in that they afford significant flexibility in the extentto which the grinding matrix may be incorporated with the biologicallyactive material into a medicament.

In a highly preferred form, the grinding matrix is harder than thebiologically active material, and is thus capable of reducing theparticle size of the active material under the dry milling conditions ofthe invention. Again without wishing to be bound by theory, under thesecircumstances it is believed that the millable grinding matrix affordsthe advantage of the present invention through a second route, with thesmaller particles of grinding matrix produced under the dry millingconditions enabling greater interaction with the biologically activematerial. The quantity of the grinding matrix relative to the quantityof biologically active material, and the extent of physical degradationof the grinding matrix, is sufficient to improve to inhibitre-agglomeration of the particles of the active material. Preferably,the quantity of the grinding matrix relative to the quantity ofbiologically active material, and the extent of physical degradation ofthe grinding matrix, is sufficient to inhibit re-agglomeration of theparticles of the active material in nanoparticulate form.

The grinding matrix is not generally selected to be chemically reactivewith the biologically active material under the milling conditions ofthe invention, excepting for example, where the matrix is deliberatelychosen to undergo a mechanico-chemical reaction. Such a reaction mightbe the conversion of a free base or acid to a salt or vice versa.

As stated above, the method of the present invention requires thegrinding matrix to be milled with the biologically active material; thatis, the grinding matrix will physically degrade under the dry millingconditions of the invention to facilitate the formation and retention ofparticulates of the biologically active material with reduced particlesize. The precise extent of degradation required will depend on certainproperties of the grinding matrix and the biologically active material,the ratio of biologically active material to grinding matrix, and theparticle size distribution of the particles comprising the biologicallyactive material.

The physical properties of the grinding matrix necessary to achieve therequisite degradation are dependent on the precise milling conditions.For example, a harder grinding matrix may degrade to a sufficient extentprovided it is subjected to more vigorous dry milling conditions.Physical properties of the grinding matrix relevant to the extent thatthe agent will degrade under dry milling conditions include hardness,friability, as measured by indicia such as hardness, fracture toughnessand brittleness index.

A low hardness (typically a Mohs Hardness less than 7) of thebiologically active material is desirable to ensure fracture of theparticles during processing, so that composite microstructures developduring milling. Preferably, the hardness is less than 3 as determinedusing the Mohs Hardness scale.

Preferably, the grinding matrix is of low abrasivity. Low abrasivity isdesirable to minimise contamination of the mixture of the biologicallyactive material in the grinding matrix by the milling bodies and/or themilling chamber of the media mill. An indirect indication of theabrasivity can be obtained by measuring the level of milling-basedcontaminants.

Preferably, the grinding matrix has a low tendency to agglomerate duringdry milling. While it is difficult to objectively quantify the tendencyto agglomerate during milling, it is possible to obtain a subjectivemeasure by observing the level of “caking” of the grinding matrix on themilling bodies and the milling chamber of the media mill as dry millingprogresses.

The grinding matrix may be an inorganic or organic substance.

In one embodiment, the grinding matrix is selected from the following,either as a single substance or a combination of two or more substances:Polyols (sugar alcohols) for example (but not limited to) mannitol,sorbitol, isomalt, xylitol, maltitol, lactitol, erythritol, arabitol,ribitol, monosaccharides for example (but not limited to) glucose,fructose, mannose, galactose, disaccharides and trisaccharides forexample (but not limited to) anhydrous lactose, lactose monohydrate,sucrose, maltose, trehalose, polysaccharides for example (but notlimited to) maltodextrins, dextrin, Inulin, dextrates, polydextrose,other carbohyrates for example (but not limited to) starch, wheat flour,corn flour, rice flour, rice starch, tapioca flour, tapioca starch,potato flour, potato starch, other flours and starches, soy flour, soymeal or other soy products, cellulose, microcrystalline cellulose,microcrystalline cellulose based co blended excipients, chemicallymodified excipients such as pregelatinized (or partially) starch,modified celluloses such as HPMC, CMC, HPC, enteric polymer coatingssuch as hypromellose phthalate, cellulose acetate phthalate (Aquacoat®),polyvinyl acetate phthalate (Sureteric®), hypromellose acetate succinate(AQOAT®), and polmethacrylates (Eudragit® and Acryl-EZE®), Milk productsfor example (but not limited to) milk powder, skim milk powders, othermilk solids and dreviatives, other functional Excipients, organic acidsfor example (but not limited to) citric acid, tartaric acid, malic acid,maleic acid fumaric acid, ascorbic acid, succinic acid, the conjugatesalt of organic acids for example (but not limited to) sodium citrate,sodium tartrate, sodium malate, sodium ascorbate, potassium citrate,potassium tartrate, potassium malate, potassium ascorbate, inorganicssuch as sodium carbonate, potassium carbonate, magnesium carbonate,sodium bicarbonate, potassium bicarbonate and calcium carbonate. dibasiccalcium phosphate, tribasic calcium phosphate, sodium sulfate, sodiumchloride, sodium metabisulphite, sodium thiosulfate, ammonium chloride,Glauber's salt, ammonium carbonate, sodium bisulfate, magnesium sulfate,potash alum, potassium chloride, sodium hydrogen sulfate, sodiumhydroxide, crystalline hydroxides, hydrogen carbonates, hydrogencarbonates of pharmaceutical acceptable alkali metals, such as but notlimited by, sodium, potassium, lithium, calcium, and barium, ammoniumsalts (or salts of volatile amines), for example (but not limited to)ammonium chloride, methylamine hydrochloride, ammonium bromide, otherinorganics for example (but not limited to), thermal silica, chalk,mica, silica, alumina, titanium dioxide, talc, kaolin, bentonite,hectorite, magnesium trisilicate, other clay or clay derivatives oraluminium silicates, a surfactant for example (but not limited to)sodium lauryl sulfate, sodium stearyl sulfate, sodium cetyl sulfate,sodium cetostearyl sulfate, sodium docusate, sodium deoxycholate,N-lauroylsarcosine sodium salt, glyceryl monostearate, glyceroldistearate glyceryl palmitostearate, glyceryl behenate, glycerylcaprylate, glyceryl oleate, benzalkonium chloride, CTAB, CTAC,Cetrimide, cetylpyridinium chloride, cetylpyridinium bromide,benzethonium chloride, PEG 40 stearate, PEG 100 stearate, poloxamer 188,poloxamer 338, poloxamer 407 polyoxyl 2 stearyl ether, polyoxyl 100stearyl ether, polyoxyl 20 stearyl ether, polyoxyl 10 stearyl ether,polyoxyl 20 cetyl ether, polysorbate 20, polysorbate 40, polysorbate 60,polysorbate 61, polysorbate 65, polysorbate 80, polyoxyl 35 castor oil,polyoxyl 40 castor oil, polyoxyl 60 castor oil, polyoxyl 100 castor oil,polyoxyl 200 castor oil, polyoxyl 40 hydrogenated castor oil, polyoxyl60 hydrogenated castor oil, polyoxyl 100 hydrogenated castor oil,polyoxyl 200 hydrogenated castor oil, cetostearyl alcohol, macrogel 15hydroxystearate, sorbitan monopalmitate, sorbitan monostearate, sorbitantrioleate, Sucrose Palmitate, Sucrose Stearate, Sucrose Distearate,Sucrose laurate, Glycocholic acid, sodium Glycholate, Cholic Acid,Sodium Cholate, Sodium Deoxycholate, Deoxycholic acid, Sodiumtaurocholate, taurocholic acid, Sodium taurodeoxycholate,taurodeoxycholic acid, soy lecithin, phosphatidylcholine,phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol,PEG4000, PEG6000, PEG8000, PEG10000, PEG20000, alkyl naphthalenesulfonate condensate/Lignosulfonate blend, Calcium DodecylbenzeneSulfonate, Sodium Dodecylbenzene Sulfonate, Diisopropylnaphthaenesulphonate, erythritol distearate, Naphthalene SulfonateFormaldehyde Condensate, nonylphenol ethoxylate (poe-30),Tristyrylphenol Ethoxylate, Polyoxyethylene (15) tallowalkylamines,sodium alkyl naphthalene sulfonate, sodium alkyl naphthalene sulfonatecondensate, sodium alkylbenzene sulfonate, sodium isopropyl naphthalenesulfonate, Sodium Methyl Naphthalene Formaldehyde Sulfonate, sodiumn-butyl naphthalene sulfonate, tridecyl alcohol ethoxylate (poe-18),Triethanolamine isodecanol phosphate ester, Triethanolaminetristyrylphosphate ester, Tristyrylphenol Ethoxylate Sulfate,Bis(2-hydroxyethyl)tallowalkylamines.

In a preferred embodiment, the grinding matrix is a matrix that isconsidered GRAS (generally regarded as safe) by persons skilled in thepharmaceutical arts.

In another preferred aspect a combination of two or more suitablematrices, such as those listed above, can be used as the grinding matrixto provide improved properties such as the reduction of caking, andgreater improvement of particle size reduction. Combination matrices mayalso be advantageous when the matrices have different solubility'sallowing the removal or partial removal of one matrix, while leaving theother or part of the other to provide encapsulation or partialencapsulation of the biologically active material.

Another highly preferred aspect of the method is the inclusion of asuitable milling aid in the matrix to improve milling performance.Improvements to milling performance would be things such as, but notlimited to, a reduction in caking or higher recovery of powder from themill. Examples of suitable milling aids include surfactants, polymersand inorganics such as silica (including colloidal silica), aluminiumsilicates and clays.

There are a wide range of surfactants that will make suitable millingaids. The highly preferred form is where the surfactant is a solid, orcan be manufactured into a solid. Preferably, the surfactant is selectedfrom the group consisting of: polyoxyethylene alkyl ethers,polyoxyethylene stearates, polyethylene glycols (PEG), poloxamers,poloxamines, sarcosine based surfactants, polysorbates, aliphaticalcohols, alkyl and aryl sulfates, alkyl and aryl polyether sulfonatesand other sulfate surfactants, trimethyl ammonium based surfactants,lecithin and other phospholipids, bile salts, polyoxyethylene castor oilderivatives, polyoxyethylene sorbitan fatty acid esters, Sorbitan fattyacid esters, Sucrose fatty acid esters, alkyl glucopyranosides, alkylmaltopyranosides, glycerol fatty acid esters, Alkyl Benzene SulphonicAcids, Alkyl Ether Carboxylic Acids, Alkyl and aryl Phosphate esters,Alkyl and aryl Sulphate esters, Alkyl and aryl Sulphonic acids, AlkylPhenol Phosphates esters, Alkyl Phenol Sulphates esters, Alkyl and ArylPhosphates, Alkyl Polysaccharides, Alkylamine Ethoxylates,Alkyl-Naphthalene Sulphonates formaldehyde condensates, Sulfosuccinates,lignosulfonates, Ceto-Oleyl Alcohol Ethoxylates, Condensed NaphthaleneSulphonates, Dialkyl and Alkyl Naphthalene Sulphonates, Di-alkylSulphosuccinates, Ethoxylated nonylphenols, Ethylene Glycol Esters,Fatty Alcohol Alkoxylates, Hydrogenated tallowalkylamines, Mono-alkylSulphosuccinamates, Nonyl Phenol Ethoxylates, Sodium Oleyl N-methylTaurate, Tallowalkylamines, linear and branched dodecylbenzene sulfonicacidsPreferably, the surfactant is selected from the group consistingof: sodium lauryl sulfate, sodium stearyl sulfate, sodium cetyl sulfate,sodium cetostearyl sulfate, sodium docusate, sodium deoxycholate,N-lauroylsarcosine sodium salt, glyceryl monostearate, glyceroldistearate glyceryl palmitostearate, glyceryl behenate, glycerylcaprylate, glyceryl oleate, benzalkonium chloride, CTAB, CTAC,Cetrimide, cetylpyridinium chloride, cetylpyridinium bromide,benzethonium chloride, PEG 40 stearate, PEG 100 stearate, poloxamer 188,poloxamer 338, poloxamer 407 polyoxyl 2 stearyl ether, polyoxyl 100stearyl ether, polyoxyl 20 stearyl ether, polyoxyl 10 stearyl ether,polyoxyl 20 cetyl ether, polysorbate 20, polysorbate 40, polysorbate 60,polysorbate 61, polysorbate 65, polysorbate 80, polyoxyl 35 castor oil,polyoxyl 40 castor oil, polyoxyl 60 castor oil, polyoxyl 100 castor oil,polyoxyl 200 castor oil, polyoxyl 40 hydrogenated castor oil, polyoxyl60 hydrogenated castor oil, polyoxyl 100 hydrogenated castor oil,polyoxyl 200 hydrogenated castor oil, cetostearyl alcohol, macrogel 15hydroxystearate, sorbitan monopalmitate, sorbitan monostearate, sorbitantrioleate, Sucrose Palmitate, Sucrose Stearate, Sucrose Distearate,Sucrose laurate, Glycocholic acid, sodium Glycholate, Cholic Acid,Sodium Cholate, Sodium Deoxycholate, Deoxycholic acid, Sodiumtaurocholate, taurocholic acid, Sodium taurodeoxycholate,taurodeoxycholic acid, soy lecithin, phosphatidylcholine,phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol,PEG4000, PEG6000, PEG8000, PEG10000, PEG20000, alkyl naphthalenesulfonate condensate/Lignosulfonate blend, Calcium DodecylbenzeneSulfonate, Sodium Dodecylbenzene Sulfonate, Diisopropylnaphthaenesulphonate, erythritol distearate, Naphthalene SulfonateFormaldehyde Condensate, nonylphenol ethoxylate (poe-30),Tristyrylphenol Ethoxylate, Polyoxyethylene (15) tallowalkylamines,sodium alkyl naphthalene sulfonate, sodium alkyl naphthalene sulfonatecondensate, sodium alkylbenzene sulfonate, sodium isopropyl naphthalenesulfonate, Sodium Methyl Naphthalene Formaldehyde Sulfonate, sodiumn-butyl naphthalene sulfonate, tridecyl alcohol ethoxylate (poe-18),Triethanolamine isodecanol phosphate ester, Triethanolaminetristyrylphosphate ester, Tristyrylphenol Ethoxylate Sulfate,Bis(2-hydroxyethyl)tallowalkylamines.

Preferably the polymer is selected from the list of:polyvinylpyrrolidones (PVP), polyvinylalcohol, Acrylic acid basedpolymers and copolymers of acrylic acid

Preferably, the milling aid has a concentration selected from the groupconsisting of: 0.1-10% w/w, 0.1-5% w/w, 0.1-2.5% w/w, of 0.1-2% w/w,0.1-1%, 0.5-5% w/w, 0.5-3% w/w, 0.5-2% w/w, 0.5-1.5%, 0.5-1 w/w, of0.75-1.25% w/w, 0.75-1% and 1% w/w.

Milling Bodies

In the method of the present invention, the milling bodies arepreferably chemically inert and rigid. The term “chemically-inert”, asused herein, means that the milling bodies do not react chemically withthe biologically active material or the grinding matrix.

As described above, the milling bodies are essentially resistant tofracture and erosion in the milling process.

The milling bodies are desirably provided in the form of bodies whichmay have any of a variety of smooth, regular shapes, flat or curvedsurfaces, and lacking sharp or raised edges. For example, suitablemilling bodies can be in the form of bodies having ellipsoidal, ovoid,spherical or right cylindrical shapes. Preferably, the milling bodiesare provided in the form of one or more of beads, balls, spheres, rods,right cylinders, drums or radius-end right cylinders (i.e., rightcylinders having hemispherical bases with the same radius as thecylinder).

Depending on the nature of the biologically active material and thegrinding matrix, the milling media bodies desirably have an effectivemean particle diameter (i.e. “particle size”) between about 0.1 and 30mm, more preferably between about 1 and about 15 mm, still morepreferably between about 3 and 10 mm.

The milling bodies may comprise various substances such as ceramic,glass, metal or polymeric compositions, in a particulate form. Suitablemetal milling bodies are typically spherical and generally have goodhardness (i.e. RHC 60-70), roundness, high wear resistance, and narrowsize distribution and can include, for example, balls fabricated fromtype 52100 chrome steel, type 316 or 440C stainless steel or type 1065high carbon steel.

Preferred ceramics, for example, can be selected from a wide array ofceramics desirably having sufficient hardness and resistance to fractureto enable them to avoid being chipped or crushed during milling and alsohaving sufficiently high density. Suitable densities for milling mediacan range from about 1 to 15 g/cm³′, preferably from about 1 to 8 g/cm³.Preferred ceramics can be selected from steatite, aluminum oxide,zirconium oxide, zirconia-silica, yttria-stabilized zirconium oxide,magnesia-stabilized zirconium oxide, silicon nitride, silicon carbide,cobalt-stabilized tungsten carbide, and the like, as well as mixturesthereof.

Preferred glass milling media are spherical (e.g. beads), have a narrowsize distribution, are durable, and include, for example, lead-free sodalime glass and borosilicate glass. Polymeric milling media arepreferably substantially spherical and can be selected from a wide arrayof polymeric resins having sufficient hardness and friability to enablethem to avoid being chipped or crushed during milling,abrasion-resistance to minimize attrition resulting in contamination ofthe product, and freedom from impurities such as metals, solvents, andresidual monomers. Preferred polymeric resins, for example, can beselected from crosslinked polystyrenes, such as polystyrene crosslinkedwith divinylbenzene, styrene copolymers, polyacrylates such aspolymethylmethacrylate, polycarbonates, polyacetals, vinyl chloridepolymers and copolymers, polyurethanes, polyamides, high densitypolyethylenes, polypropylenes, and the like. The use of polymericmilling media to grind materials down to a very small particle size (asopposed to mechanochemical synthesis) is disclosed, for example, in U.S.Pat. Nos. 5,478,705 and 5,500,331. Polymeric resins typically can havedensities ranging from about 0.8 to 3.0 g/cm³. Higher density polymericresins are preferred. Alternatively, the milling media can be compositeparticles comprising dense core particles having a polymeric resinadhered thereon. Core particles can be selected from substances known tobe useful as milling media, for example, glass, alumina, zirconiasilica, zirconium oxide, stainless steel, and the like. Preferred coresubstances have densities greater than about 2.5 g/cm³.

In one embodiment of the invention, the milling media are formed from aferromagnetic substance, thereby facilitating removal of contaminantsarising from wear of the milling media by the use of magnetic separationtechniques.

Each type of milling body has its own advantages. For example, metalshave the highest specific gravities, which increase grinding efficiencydue to increased impact energy. Metal costs range from low to high, butmetal contamination of final product can be an issue. Glasses areadvantageous from the standpoint of low cost and the availability ofsmall bead sizes as low as 0.004 mm. However, the specific gravity ofglasses is lower than other media and significantly more milling time isrequired. Finally, ceramics are advantageous from the standpoint of lowwear and contamination, ease of cleaning, and high hardness.

Dry Milling

In the dry milling process of the present invention, the biologicallyactive material and grinding matrix, in the form of crystals, powders,or the like, are combined in suitable proportions with the plurality ofmilling bodies in a milling chamber that is mechanically agitated (i.e.with or without stirring) for a predetermined period of time at apredetermined intensity of agitation. Typically, a milling apparatus isused to impart motion to the milling bodies by the external applicationof agitation, whereby various translational, rotational or inversionmotions or combinations thereof are applied to the milling chamber andits contents, or by the internal application of agitation through arotating shaft terminating in a blade, propeller, impeller or paddle orby a combination of both actions.

During milling, motion imparted to the milling bodies can result inapplication of shearing forces as well as multiple impacts or collisionshaving significant intensity between milling bodies and particles of thebiologically active material and grinding matrix. The nature andintensity of the forces applied by the milling bodies to thebiologically active material and the grinding matrix is influenced by awide variety of processing parameters including: the type of millingapparatus; the intensity of the forces generated, the kinematic aspectsof the process; the size, density, shape, and composition of the millingbodies; the weight ratio of the biologically active material andgrinding matrix mixture to the milling bodies; the duration of milling;the physical properties of both the biologically active material and thegrinding matrix; the atmosphere present during activation; and others.

Advantageously, the media mill is capable of repeatedly or continuouslyapplying mechanical compressive forces and shear stress to thebiologically active material and the grinding matrix. Suitable mediamills include but are not limited to the following: high-energy ball,sand, bead or pearl mills, basket mill, planetary mill, vibratory actionball mill, multi-axial shaker/mixer, stirred ball mill, horizontal smallmedia mill, multi-ring pulverizing mill, and the like, including smallmilling media. The milling apparatus also can contain one or morerotating shafts.

In a preferred form of the invention, the dry milling is performed in aball mill. Throughout the remainder of the specification reference willbe made to dry milling being carried out by way of a ball mill. Examplesof this type of mill are attritor mills, nutating mills, tower mills,planetary mills, vibratory mills and gravity-dependent-type ball mills.It will be appreciated that dry milling in accordance with the method ofthe invention may also be achieved by any suitable means other than ballmilling. For example, dry milling may also be achieved using jet mills,rod mills, roller mills or crusher mills.

Biologically Active Material

The biologically active material includes active compounds, includingcompounds for veterinary and human use such as but not limited to,pharmaceutical actives and the like.

The biologically active material is ordinarily a material for which oneof skill in the art desires improved dissolution properties. Thebiologically active material may be a conventional active agent or drug,although the process of the invention may be employed on formulations oragents that already have reduced particle size compared to theirconventional form.

Biologically active materials suitable for use in the invention includenaproxen.

As discussed in the context of the background to the invention,biologically active materials that are poorly water soluble atgastrointestinal pH will particularly benefit from being prepared, andthe method of the present invention is particularly advantageouslyapplied to materials that are poorly water soluble at gastrointestinalpH.

Conveniently, the biologically active material is capable ofwithstanding temperatures that are typical in uncooled dry milling,which may exceed 80° C. Therefore, materials with a melting point about80° C. or greater are highly suitable. For biologically active materialswith lower melting points, the media mill may be cooled, therebyallowing materials with significantly lower melting temperatures to beprocessed according to the method of the invention. For instance, asimple water-cooled mill will keep temperatures below 50° C., or chilledwater could be used to further lower the milling temperature. Thoseskilled in the art will understand that a high energy ball mill could bedesigned to run at any temperature between say −30 to 200° C. For somebiologically active materials it may be advantageous to control themilling temperature to temperatures significantly below the meltingpoints of the biologically active materials.

The biologically active material is obtained in a conventional formcommercially and/or prepared by techniques known in the art.

It is preferred, but not essential, that the particle size of thebiologically active material be less than about 1000 μm, as determinedby sieve analysis. If the coarse particle size of the biologicallyactive material is greater than about 1000 μm, then it is preferred thatthe particles of the biologically active material substrate be reducedin size to less than 1000 μm using another standard milling method.

Processed Biologically Active Material

Preferably, the biologically active materials, which have been subjectto the methods of the invention, comprises particles of biologicallyactive material of an average particle size, determined on a particlenumber basis, is equal to or less than a size selected from the group2000 nm, 1900 nm, 1800 nm, 1700 nm, 1600 nm, 1500 nm, 1400 nm, 1300 nm,1200 nm, 1100 nm, 1000 nm, 900 nm, 800 nm, 700 nm, 600 nm, 500 nm, 400nm, 300 nm, 200 nm and 100 nm.

Preferably, the biologically active materials, which have been subjectto the methods of the invention, comprises particles of biologicallyactive material of a median particle size, determined on a particlevolume basis, equal or less than a size selected from the group 2000 nm,1900 nm, 1800 nm, 1700 nm, 1600 nm, 1500 nm, 1400 nm, 1300 nm, 1200 nm,1100 nm, 1000 nm, 900 nm, 800 nm, 700 nm, 600 nm, 500 nm, 400 nm, 300nm, 200 nm and 100 nm.

Preferably, the biologically active materials, which have been subjectto the methods of the invention, comprises particles of biologicallyactive material and wherein the Dx of the particle size distribution, asmeasured on a particle volume basis, is selected from the groupconsisting of less than or equal to 10,000 nm, 5000 nm, 3000 nm, 2000nm, 1900 nm, 1800 nm, 1700 nm, 1600 nm, 1500 nm, 1400 nm, 1300 nm, 1200nm, 1100 nm, 1000 nm, 900 nm, 800 nm, 700 nm, 600 nm, 500 nm, 400 nm,300 nm, 200 nm, and 100 nm; wherein x is greater than or equal to 90,These sizes refer to particles either fully dispersed or partiallyagglomerated.

Agglomerates of Biologically Active Material after Processing

Agglomerates comprising particles of biologically active material, saidparticles having a particle size within the ranges specified above,should be understood to fall within the scope of the present invention,regardless of whether the agglomerates exceed the ranges specifiedabove.

Agglomerates comprising particles of biologically active material, saidagglomerates having a total agglomerate size within the ranges specifiedabove, should be understood to fall within the scope of the presentinvention.

Agglomerates comprising particles of biologically active material,should be understood to fall within the scope of the present inventionif at the time of use, or further processing, the particle size of theagglomerate is within the ranges specified above.

Agglomerates comprising particles of biologically active material, saidparticles having a particle size within the ranges specified above, atthe time of use, or further processing, should be understood to fallwithin the scope of the present invention, regardless of whether theagglomerates exceed the ranges specified above.

Processing Time

Preferably, the biologically active material and the grinding matrix aredry milled for the shortest time necessary to form the mixture of thebiologically active material in the grinding matrix such that the activematerial has improved dissolution to minimise any possible contaminationfrom the media mill and/or the plurality of milling bodies. This timevaries greatly, depending on the biologically active material and thegrinding matrix, and may range from as short as 1 minute to severalhours. Dry milling times in excess of 2 hours may lead to degradation ofthe biologically active material and an increased level of undesirablecontaminants.

Suitable rates of agitation and total milling times are adjusted for thetype and size of milling apparatus as well as the milling media, theweight ratio of the biologically active material and grinding matrixmixture to the plurality of milling bodies, the chemical and physicalproperties of the biologically active material and grinding matrix, andother parameters that may be optimized empirically.

Inclusion of the Grinding Matrix with the Biologically Active Materialand Separation of the Grinding Matrix from the Biologically ActiveMaterial

In a preferred aspect, the grinding matrix is not separated from thebiologically active material but is maintained with the biologicallyactive material in the final product. Preferably the grinding matrix isconsidered to be Generally Regarded as Safe (GRAS) for pharmaceuticalproducts. In an alternative aspect, the grinding matrix is separatedfrom the biologically active material. In one aspect, where the grindingmatrix is not fully milled, the unmilled grinding matrix is separatedfrom the biologically active material. In a further aspect, at least aportion of the milled grinding matrix is separated from the biologicallyactive material.

Any portion of the grinding matrix may be removed, including but notlimited to 10%, 25%, 50%, 75%, or substantially all of the grindingmatrix.

In some embodiments of the invention, a significant portion of themilled grinding matrix may comprise particles of a size similar toand/or smaller than the particles comprising the biologically activematerial. Where the portion of the milled grinding matrix to beseparated from the particles comprising the biologically active materialcomprises particles of a size similar to and/or smaller than theparticles comprising the biologically active material, separationtechniques based on size distribution are inapplicable.

In these circumstances, the method of the present invention may involveseparation of at least a portion of the milled grinding matrix from thebiologically active material by techniques including but not limited toelectrostatic separation, magnetic separation, centrifugation (densityseparation), hydrodynamic separation, froth flotation.

Advantageously, the step of removing at least a portion of the milledgrinding matrix from the biologically active material may be performedthrough means such as selective dissolution, washing, or sublimation.

An advantageous aspect of the invention would be the use of grindingmatrix that has two or more components where at least one component iswater soluble and at least one component has low solubility in water. Inthis case washing can be used to remove the matrix component soluble inwater leaving the biologically active material encapsulated in theremaining matrix components. In a highly advantageous aspect of theinvention the matrix with low solubility is a functional excipient.

A highly advantageous aspect of the present invention is that certaingrinding matrixes appropriate for use in the method of the invention (inthat they physically degrade to the desired extent under dry millingconditions) are also pharmaceutically acceptable and thus appropriatefor use in a medicament. Where the method of the present invention doesnot involve complete separation of the grinding matrix from thebiologically active material, the present invention encompasses methodsfor the production of a medicament incorporating both the biologicallyactive material and at least a portion of the milled grinding matrix,medicaments so produced and methods of treatment of an animal, includingman, using a therapeutically effective amount of said biologicallyactive materials by way of said medicaments.

The medicament may include only the biologically active material and thegrinding matrix or, more preferably, the biologically active materialsand grinding matrix may be combined with one or more pharmaceuticallyacceptable carriers, as well as any desired excipients or other likeagents commonly used in the preparation of medicaments.

Analogously, a highly advantageous aspect of the present invention isthat certain grinding matrixes appropriate for use in the method of theinvention (in that they physically degrade to a desirable extent underdry milling conditions) are also appropriate for use in an agriculturalchemical composition. Where the method of the present invention does notinvolve complete separation of the grinding matrix from the biologicallyactive material, the present invention encompasses methods for theproduction of a agricultural chemical composition incorporating both thebiologically active material and at least a portion of the milledgrinding matrix, agricultural chemical composition so produced andmethods of use of such compositions.

The agricultural chemical composition may include only the biologicallyactive material and the grinding matrix or, more preferably, thebiologically active materials and grinding matrix may be combined withone or more acceptable carriers, as well as any desired excipients orother like agents commonly used in the preparation of agriculturalchemical compositions.

In one particular form of the invention, the grinding matrix is bothappropriate for use in a medicament and readily separable from thebiologically active material by methods not dependent on particle size.Such grinding matrixes are described in the following detaileddescription of the invention. Such grinding matrixes are highlyadvantageous in that they afford significant flexibility in the extentto which the grinding matrix may be incorporated with the biologicallyactive material into a medicament.

The mixture of biologically active material and grinding matrix may thenbe separated from the milling bodies and removed from the mill.

In one embodiment, the grinding matrix is separated from the mixture ofbiologically active material and grinding matrix. Where the grindingmatrix is not fully milled, the unmilled grinding matrix is separatedfrom the biologically active material. In a further aspect, at least aportion of the milled grinding matrix is separated from the biologicallyactive material.

The milling bodies are essentially resistant to fracture and erosion inthe dry milling process. The quantity of the grinding matrix relative tothe quantity of biologically active material, and the extent of millingof the grinding matrix, is sufficient to provide reduced particle sizeof the biologically active material.

The grinding matrix is neither chemically nor mechanically reactive withthe pharmaceutical material under the dry milling conditions of themethod of the invention except, for example, where the matrix isdeliberately chosen to undergo a mechanico-chemical reaction. Such areaction might be the conversion of a free base or acid to a salt orvice versa.

Preferably, the medicament is a solid dosage form, however, other dosageforms may be prepared by those of ordinary skill in the art.

In one form, after the step of separating said mixture of biologicallyactive material and grinding matrix from the plurality of millingbodies, and before the step of using said mixture of biologically activematerial and grinding matrix in the manufacture of a medicament, themethod may comprise the step of:

removing a portion of the grinding matrix from said mixture ofbiologically active material and grinding matrix to provide a mixtureenriched in the biologically active material;and the step of using said mixture of biologically active material andgrinding matrix in the manufacture of a medicament, more particularlycomprises the step of using the mixture of biologically active materialand grinding matrix enriched in the biologically active material form inthe manufacture of a medicament.

The present invention includes medicaments manufactured by said methods,and methods for the treatment of an animal, including man, by theadministration of a therapeutically effective amount of the biologicallyactive materials by way of said medicaments.

In another embodiment of the invention, a facilitating agent or acombination of facilitating agents is also comprised in the mixture tobe milled. Such facilitating agents appropriate for use in the inventioninclude diluents, surfactants, polymers, binding agents, filling agents,lubricating agents, sweeteners, flavouring agents, preservatives,buffers, wetting agents, disintegrants, effervescent agents and agentsthat may form part of a medicament, including a solid dosage form, orother excipients required for other specific drug delivery, such as theagents and media listed below under the heading Medicinal andPharmaceutical Compositions, or any combination thereof.

Biologically Active Materials and Compositions

The present invention encompasses pharmaceutically acceptable materialsproduced according to the methods of the present invention, compositionsincluding such materials, including compositions comprising suchmaterials together with the grinding matrix with or without millingaids, facilitating agents, with at least a portion of the grindingmatrix or separated from the grinding matrix.

The pharmaceutically acceptable materials within the compositions of theinvention are present at a concentration of between about 0.1% and about99.0% by weight. Preferably, the concentration of pharmaceuticallyacceptable materials within the compositions will be about 5% to about80% by weight, while concentrations of 10% to about 50% by weight arehighly preferred. Desirably, the concentration will be in the range ofabout 10 to 15% by weight, 15 to 20% by weight, 20 to 25% by weight, 25to 30% by weight, 30 to 35% by weight, 35 to 40% by weight, 40 to 45% byweight, 45 to 50% by weight, 50 to 55% by weight, 55 to 60% by weight,60 to 65% by weight, 65 to 70% by weight, 70 to 75% by weight or 75 to80% by weight for the composition prior to any later removal (ifdesired) of any portion of the grinding matrix. Where part or the entiregrinding matrix has been removed, the relative concentration ofpharmaceutically acceptable materials in the composition may beconsiderably higher depending on the amount of the grinding matrix thatis removed. For example, if the entire grinding matrix is removed theconcentration of particles in the preparation may approach 100% byweight (subject to the presence of facilitating agents).

Compositions produced according to the present invention are not limitedto the inclusion of a single species of pharmaceutically acceptablematerials. More than one species of pharmaceutically acceptablematerials may therefore be present in the composition. Where more thanone species of pharmaceutically acceptable materials is present, thecomposition so formed may either be prepared in a dry milling step, orthe pharmaceutically acceptable materials may be prepared separately andthen combined to form a single composition.

Medicaments

The medicaments of the present invention may include thepharmaceutically acceptable material, optionally together with thegrinding matrix or at least a portion of the grinding matrix, with orwithout milling aids, facilitating agents, combined with one or morepharmaceutically acceptable carriers, as well as other agents commonlyused in the preparation of pharmaceutically acceptable compositions.

As used herein “pharmaceutically acceptable carrier” includes any andall solvents, dispersion media, coatings, antibacterial and antifungalagents, isotonic and absorption delaying agents, and the like that arephysiologically compatible. Preferably, the carrier is suitable forparenteral administration, intravenous, intraperitoneal, intramuscular,sublingual, pulmonary, transdermal or oral administration.Pharmaceutically acceptable carriers include sterile aqueous solutionsor dispersions and sterile powders for the extemporaneous preparation ofsterile injectable solutions or dispersion. The use of such media andagents for the manufacture of medicaments is well known in the art.Except insofar as any conventional media or agent is incompatible withthe pharmaceutically acceptable material, use thereof in the manufactureof a pharmaceutical composition according to the invention iscontemplated.

Pharmaceutical acceptable carriers according to the invention mayinclude one or more of the following examples:

-   -   (1) surfactants and polymers, including, but not limited to        polyethylene glycol (PEG), polyvinylpyrrolidone (PVP),        polyvinylalcohol, crospovidone,        polyvinylpyrrolidone-polyvinylacrylate copolymer, cellulose        derivatives, hydroxypropylmethyl cellulose, hydroxypropyl        cellulose, carboxymethylethyl cellulose, hydroxypropyllmethyl        cellulose phthalate, polyacrylates and polymethacrylates, urea,        sugars, polyols, and their polymers, emulsifiers, sugar gum,        starch, organic acids and their salts, vinyl pyrrolidone and        vinyl acetate; and or    -   (2) binding agents such as various celluloses and cross-linked        polyvinylpyrrolidone, microcrystalline cellulose; and or    -   (3) filling agents such as lactose monohydrate, lactose        anhydrous, microcrystalline cellulose and various starches; and        or    -   (4) lubricating agents such as agents that act on the        flowability of the powder to be compressed, including colloidal        silicon dioxide, talc, stearic acid, magnesium stearate, calcium        stearate, silica gel; and or    -   (5) sweeteners such as any natural or artificial sweetener        including sucrose, xylitol, sodium saccharin, cyclamate,        aspartame, and accsulfame K; and or    -   (6) flavouring agents; and or    -   (7) preservatives such as potassium sorbate, methylparaben,        propylparaben, benzoic acid and its salts, other esters of        parahydroxybenzoic acid such as butylparaben, alcohols such as        ethyl or benzyl alcohol, phenolic chemicals such as phenol, or        quarternary compounds such as benzalkonium chloride; and or    -   (8) buffers; and or    -   (9) Diluents such as pharmaceutically acceptable inert fillers,        such as microcrystalline cellulose, lactose, dibasic calcium        phosphate, saccharides, and/or mixtures of any of the foregoing;        and or    -   (10) wetting agents such as corn starch, potato starch, maize        starch, and modified starches, croscarmellose sodium,        crosspovidone, sodium starch glycolate, and mixtures thereof;        and or    -   (11) disintegrants; and or    -   (12) effervescent agents such as effervescent couples such as an        organic acid (e.g., citric, tartaric, malic, fumaric, adipic,        succinic, and alginic acids and anhydrides and acid salts), or a        carbonate (e.g. sodium carbonate, potassium carbonate, magnesium        carbonate, sodium glycine carbonate, L-lysine carbonate, and        arginine carbonate) or bicarbonate (e.g. sodium bicarbonate or        potassium bicarbonate); and or    -   (13) other pharmaceutically acceptable excipients.

Medicaments of the invention suitable for use in animals and inparticular in man typically must be stable under the conditions ofmanufacture and storage. The medicaments of the invention comprising thebiologically active material can be formulated as a solid, a solution, amicroemulsion, a liposome, or other ordered structures suitable to highdrug concentration. Actual dosage levels of the biologically activematerial in the medicament of the invention may be varied in accordancewith the nature of the biologically active material, as well as thepotential increased efficacy due to the advantages of providing andadministering the biologically active material (e.g., increasedsolubility, more rapid dissolution, increased surface area of thebiologically active material, etc.). Thus as used herein“therapeutically effective amount” will refer to an amount ofbiologically active material required to effect a therapeutic responsein an animal. Amounts effective for such a use will depend on: thedesired therapeutic effect; the route of administration; the potency ofthe biologically active material; the desired duration of treatment; thestage and severity of the disease being treated; the weight and generalstate of health of the patient; and the judgment of the prescribingphysician.

In another embodiment, the biologically active material, optionallytogether with the grinding matrix or at least a portion of the grindingmatrix, of the invention may be combined into a medicament with anotherbiologically active material, or even the same biologically activematerial. In the latter embodiment, a medicament may be achieved whichprovides for different release characteristics—early release from thebiologically active material, and later release from a larger averagesize biologically active material.

Pharmacokinetic Properties of Naproxen Compositions

Suitable animal models to determine pharmacokinetic parameters aredescribed in the prior art, such as the beagle dog model described inU.S. Pat. No. 7,101,576.

Fast Onset of Activity

The naproxen compositions of the invention exhibit faster therapeuticeffects.

In one example, following administration the naproxen compositions ofthe invention have a T_(max) of less than about 5 hours, less than about4.5 hours, less than about 4 hours, less than about 3.5 hours, less thanabout 3 hours, less than about 2.75 hours, less than about 2.5 hours,less than about 2.25 hours, less than about 2 hours, less than about1.75 hours, less than about 1.5 hours, less than about 1.25 hours, lessthan about 1.0 hours, less than about 50 minutes, less than about 40minutes, less than about 30 minutes, less than about 25 minutes, lessthan about 20 minutes, less than about 15 minutes, less than about 10minutes, less than about 5 minutes, or less than about 1 minute.

Increased Bioavailability

The naproxen compositions of the invention preferably exhibit increasedbioavailability (AUC) and require smaller doses as compared to priorconventional compositions administered at the same dose. Any drugcomposition can have adverse side effects. Thus, lower doses of drugswhich can achieve the same or better therapeutic effects as thoseobserved with larger doses of conventional compositions are desired.Such lower doses can be realized with the compositions of the inventionbecause the greater bioavailability observed with the compositions ascompared to conventional drug formulations means that smaller doses ofdrug are required to obtain the desired therapeutic effect.

The Pharmacokinetic Profiles of the Compositions of the Invention arenot Substantially Affected by the Fed or Fasted State of the SubjectIngesting the Compositions

The invention encompasses naproxen compositions wherein thepharmacokinetic profile of the composition is not substantially affectedby the fed or fasted state of a subject ingesting the composition. Thismeans that there is no substantial difference in the quantity ofcomposition or the rate of composition absorption when the compositionsare administered in the fed versus the fasted state. Thus, thecompositions of the invention substantially eliminate the effect of foodon the pharmacokinetics of the composition.

The difference in absorption of the naproxen composition of theinvention, when administered in the fed versus the fasted state, is lessthan about 35%, less than about 30%, less than about 25%, less thanabout 20%, less than about 15%, less than about 10%, less than about 5%,or less than about 3%. This is an especially important feature intreating patients with difficulty in maintaining a fed state.

In addition, preferably the difference in the rate of absorption (i.e.,T_(max)) of the naproxen compositions of the invention, whenadministered in the fed versus the fasted state, is less than about100%, less than about 90%, less than about 80%, less than about 70%,less than about 60%, less than about 50%, less than about 40%, less thanabout 30%, less than about 20%, less than about 15%, less than about10%, less than about 5%, less than about 3%, or essentially nodifference. Benefits of a dosage form which substantially eliminates theeffect of food include an increase in subject convenience, therebyincreasing subject compliance, as the subject does not need to ensurethat they are taking a dose either with or without food.

Preferably, the T_(max) of an administered dose of a naproxencomposition of the invention is less than that of a conventional drugactive composition, administered at the same dosage.

A preferred naproxen composition of the invention exhibits incomparative pharmacokinetic testing with a standard conventional drugactive composition, in oral suspension, capsule or tablet form, aT_(max) which is less than about 100%, less than about 90%, less thanabout 80%, less than about 70%, less than about 60%, less than about50%, less than about 40%, less than about 30%, less than about 25%, lessthan about 20%, less than about 15%, or less than about 10% of theT_(max) exhibited by the standard conventional drug active composition.

In addition, preferably the C_(max) of a naproxen composition of theinvention is greater than the C_(max) of a conventional drug activecomposition, administered at the same dosage. A preferred composition ofthe invention exhibits in comparative pharmacokinetic testing with astandard conventional drug active composition, in oral suspension,capsule or tablet form, a C_(max) which is greater than about 5%,greater than about 10%, greater than about 15%, greater than about 20%,greater than about 30%, greater than about 40%, greater than about 50%,greater than about 60%, greater than about 70%, greater than about 80%,greater than about 90%, greater than about 100%, greater than about110%, greater than about 120%, greater than about 130%, greater thanabout 140%, or greater than about 150% than the C_(max) exhibited by thestandard conventional drug active composition.

In addition, preferably the naproxen composition has an AUC greater thanthat of the equivalent conventional composition administered at the samedosage. A preferred composition of the invention exhibits in comparativepharmacokinetic testing with a standard conventional drug activecomposition, in oral suspension, capsule or tablet form, a AUC which isgreater than about 5%, greater than about 10%, greater than about 15%,greater than about 20%, greater than about 30%, greater than about 40%,greater than about 50%, greater than about 60%, greater than about 70%,greater than about 80%, greater than about 90%, greater than about 100%,greater than about 110%, greater than about 120%, greater than about130%, greater than about 140%, or greater than about 150% than the AUCexhibited by the standard conventional drug active composition.

Any standard pharmacokinetic protocol can be used to determine bloodplasma concentration profile in humans following administration of acomposition, and thereby establish whether that composition meets thepharmacokinetic criteria set out herein. For example, a randomizedsingle-dose crossover study can be performed using a group of healthyadult human subjects. The number of subjects should be sufficient toprovide adequate control of variation in a statistical analysis, and istypically about 10 or greater, although for certain purposes a smallergroup can suffice. Each subject receives by oral administration at timezero a single dose (e.g., 300 mg) of a test formulation of composition,normally at around 8 am following an overnight fast. The subjectscontinue to fast and remain in an upright position for about 4 hoursafter administration of the composition. Blood samples are collectedfrom each subject prior to administration (e.g., 15 minutes) and atseveral intervals after administration. For the present purpose it ispreferred to take several samples within the first hour, and to sampleless frequently thereafter. Illustratively, blood samples could becollected at 15, 30, 45, 60, and 90 minutes after administration, thenevery hour from 2 to 10 hours after administration. Additional bloodsamples may also be taken later, for example at 12 and 24 hours afteradministration. If the same subjects are to be used for study of asecond test formulation, a period of at least 7 days should elapsebefore administration of the second formulation. Plasma is separatedfrom the blood samples by centrifugation and the separated plasma isanalyzed for composition by a validated high performance liquidchromatography (HPLC) or liquid chromatography mass spectrometry (LCMS)procedure. Plasma concentrations of composition referenced herein areintended to mean total concentrations including both free and boundcomposition.

Any formulation giving the desired pharmacokinetic profile is suitablefor administration according to the present methods. Exemplary types offormulations giving such profiles are liquid dispersions and solid doseforms of composition. If the liquid dispersion medium is one in whichthe composition has very low solubility, the particles are present assuspended particles. The smaller the particles the higher theprobability that the formulation will exhibit the desiredpharmacokinetic profile.

Thus, a naproxen composition of the invention, upon administration to asubject, provides improved pharmacokinetic and/or pharmacodynamicproperties compared with a standard reference indomethacin compositionas measured by at least one of speed of absorption, dosage potency,efficacy, and safety.

Modes of Administration of Medicaments Comprising Biologically ActiveMaterials

Medicaments of the invention can be administered to animals, includingman, in any pharmaceutically acceptable manner, such as orally,rectally, pulmonary, intravaginally, locally (powders, ointments ordrops), transdermal, parenteral administration, intravenous,intraperitoneal, intramuscular, sublingual or as a buccal or nasalspray.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, pellets, and granules. Further, incorporating any of thenormally employed excipients, such as those previously listed, andgenerally 5-95% of the biologically active agent, and more preferably ata concentration of 10%-75% will form a pharmaceutically acceptablenon-toxic oral composition. Medicaments of the invention may beparenterally administered as a solution of the biologically active agentsuspended in an acceptable carrier, preferably an aqueous carrier. Avariety of aqueous carriers may be used, e.g. water, buffered water,0.4% saline, 0.3% glycine, hyaluronic acid and the like. Thesecompositions may be sterilized by conventional, well known sterilizationtechniques, or may be sterile filtered. The resulting aqueous solutionsmay be packaged for use as is, or lyophilized, the lyophilizedpreparation being combined with a sterile solution prior toadministration.

For aerosol administration, medicaments of the invention are preferablysupplied along with a surfactant or polymer and propellant. Thesurfactant or polymer must, of course, be non-toxic, and preferablysoluble in the propellant. Representative of such agents are the estersor partial esters of fatty acids containing from 6 to 22 carbon atoms,such as caproic, octanoic, lauric, palmitic, stearic, linoleic,linolenic, olesteric and oleic acids with an aliphatic polyhydricalcohol or its cyclic anhydride. Mixed esters, such as mixed or naturalglycerides may be employed. The surfactant or polymer may constitute0.1%-20% by weight of the composition, preferably 0.25-5%. The balanceof the composition is ordinarily propellant. A carrier can also beincluded, as desired, as with, e.g., lecithin for intranasal delivery.

Medicaments of the invention may also be administered via liposomes,which serve to target the active agent to a particular tissue, such aslymphoid tissue, or targeted selectively to cells. Liposomes includeemulsions, foams, micelles, insoluble monolayers, liquid crystals,phospholipid dispersions, lamellar layers and the like. In thesepreparations the composite microstructure composition is incorporated aspart of a liposome, alone or in conjunction with a molecule that bindsto or with other therapeutic or immunogenic compositions.

As described above, the biologically active material can be formulatedinto a solid dosage form (e.g., for oral or suppository administration),together with the grinding matrix or at least a portion of it. In thiscase there may be little or no need to add stabilizing agents since thegrinding matrix may effectively act as a solid-state stabilizer.

However, if the biologically active material is to be utilized in aliquid suspension, the particles comprising the biologically activematerial may require further stabilization once the solid carrier hasbeen substantially removed to ensure the elimination, or at leastminimisation of particle agglomeration.

Therapeutic Uses

Therapeutic uses of the medicaments of the invention include painrelief, anti-inflammatory, migraine, asthma, and other disorders thatrequire the active agent to be administered with a high bioavailability.

One of the main areas when rapid bioavailability of a biologicallyactive material is required is in the relief of pain. The minoranalgesics, such as cyclooxygenase inhibitors (aspirin related drugs)may be prepared as medicaments according to the present invention.

Medicaments of the invention may also be used for treatment of eyedisorders. That is, the biologically active material may be formulatedfor administration on the eye as an aqueous suspension in physiologicalsaline, or a gel. In addition, the biologically active material may beprepared in a powder form for administration via the nose for rapidcentral nervous system penetration.

Treatment of cardiovascular disease may also benefit from biologicallyactive materials according to the invention, such as treatment of anginapectoris and, in particular, molsidomine may benefit from betterbioavailability.

Other therapeutic uses of the medicaments of the present inventioninclude treatment of hair loss, sexual dysfunction, or dermal treatmentof psoriasis.

The present invention will now be described with reference to thefollowing non-limiting Examples. The description of the Examples is inno way limiting on the preceding paragraphs of this specification, butis provided for exemplification of the methods and compositions of theinvention.

EXAMPLES

It will be apparent to persons skilled in the milling and pharmaceuticalarts that numerous enhancements and modifications can be made to theabove described processes without departing from the basic inventiveconcepts. For example, in some applications the biologically activematerial may be pretreated and supplied to the process in the pretreatedform. All such modifications and enhancements are considered to bewithin the scope of the present invention, the nature of which is to bedetermined from the foregoing description and the appended claims.Furthermore, the following Examples are provided for illustrativepurposes only, and are not intended to limit the scope of the processesor compositions of the invention.

The Following Materials were Used in the Examples

Active pharmaceutical ingredients were sourced from commercialsuppliers, excipients from either commercial suppliers such asSigma-Aldrich or retailers, while food ingredients were sourced fromretailers.

The Following Mills were Used for the Grinding Experiments

Spex-Type Mill:

Small scale milling experiments were conducted using a vibratory Spex8000D mixer/mill. Twelve ⅜″ stainless steel balls were used as thegrinding media. The powder charge and grinding media were loaded into ahardened steel vial with an internal volume of approximately 75 mL.Following milling, the milled material was discharged from the vial andsieved to remove grinding media.

Attritor-Type Mill:

Small scale attritor milling experiments were performed using a 1 HDUnion Process attritor mill with a 110 mL grinding chamber. The grindingmedia consisted of 330 g 5/16″ stainless steel balls. The mill wasloaded through the loading port, with dry materials added initially,followed by the grinding media. The milling process was conducted withthe jacket cooled at 10-20° C. and the shaft rotating at 500 rpm. Uponcompletion of milling, the milled material was discharged from the milland sieved to remove the grinding media.

Medium scale attritor milling experiments were performed using a 1HDUnion Process attritor mill with a 1 L grinding chamber or a 1S UnionProcess attritor mill with a 750 mL grinding chamber. The grinding mediaconsisted of 3 kg of 5/16″ stainless steel balls or 1.5 kg of ⅜″stainless steel balls for the 1S attritor. The 1HD mill was loadedthrough the loading port, with dry materials added initially, followedby the grinding media, while the grinding media was added initially,followed by the dry materials in the 1S attritor mill. The millingprocess was conducted with the jacket cooled at 10-20° C. with the shaftrotating at 350 rpm in the 1HD attritor or 550 rpm in the 1S attritor.Upon completion of milling, the milled material was discharged from themill and sieved to remove the grinding media.

Medium to large scale attritor milling experiments were performed usinga 1S Union Process attritor mill with a ½ gallon grinding chamber. Thegrinding media consisted of 7 kg of ⅜″ stainless steel balls. The millwas loaded through the loading port, with the grinding media addedinitially, followed by the dry powders. The milling process wasconducted with the jacket cooled at 18° C. and the shaft rotating at550-555 rpm. Upon completion of milling, the milled powder wasdischarged from the mill through the bottom discharge port at 77 rpm for5 min. Large scale attritor milling experiments were performed using a1S Union Process attritor mill with a 1½ gallon grinding chamber. Thegrinding media consisted of 20 kg of ⅜″ stainless steel balls. The millwas loaded through the loading port, with the grinding media addedinitially, then followed by the dry powders. The milling process wasconducted with the jacket cooled to ambient temperature and the shaftrotating at 300 rpm. Upon completion of milling, the milled powder wasdischarged from the mill through the bottom discharge port at 77 rpm for5 min. The largest scale attritor millings were done in a 30S UnionProcess mill with a 25 gallon grinding chamber (Union Process, AkronOhio, USA). The grinding media consisted of 454 kg of ⅜″ stainless steelballs. The mill was loaded through its split top lid, with the grindingmedia added initially, then followed by the dry powders (25 kg). Themilling process was conducted with the jacket cooled to 10° C. and theshaft rotating at 130 rpm. Upon completion of milling, the milled powderwas discharged from the mill through the bottom discharge port at 77 rpmfor 5 min.

Siebtechnik Mill

Medium scale milling experiments were also performed in a SiebtechnikGSM06 (Siebtechnik, GmbH, Germany) with two 1 L milling chambers. Eachchamber was filled with 2.7 kg stainless steel media with a diameter of⅜″. The media and powder were loaded with the lid off. The mill wasoperated at ambient temperature. The vibration speed was the standardmill settings. Upon completion of the milling the media was separatedfrom the powder by sieving.

Simoloyer Mill

Medium scale milling experiments were performed in a Simoloyer CM01 (ZOZGmbH, Germany) with a 2 L milling chamber. The grinding media consistedof 2.5 kg stainless steel media with a diameter of 5 mm. the media wasloaded though the loading port followed by the dry materials. Themilling vessel was cooled using water at a temperature of about 18° C.The mill speed was operated in cycle mode: at 1300 rpm for two minutesand at 500 rpm for 0.5 min and so forth. Upon completion of the millingthe media was discharged from the mill using a grated valve to retainthe grinding media.

Large scale milling experiments were performed in a Simoloyer CM100 (ZOZGmbH, Germany) with a 100 L milling chamber. The grinding mediaconsisted of 100 kg stainless steel media with a diameter of 3/16″. Thepowder charge (11 kg) was added to the milling chamber, which alreadycontained the grinding media, through a loading port. The millingchamber was cooled to 18° C. and the powder was milled for a total of 20minutes using a cycling mode equivalent to a tip speed at 1300/500 rpmfor 2/0.5 min in the CM-01 type mill. Upon completion of the milling themill was discharged by sucking the powder into a cyclone.

Hicom Mill

Millings performed in a nutating Hicom mill utilized 14 kg of stainlesssteel 0.25″ grinding media together with a powder charge of 480 g. Themill was loaded by pre-mixing media and powder, then adding the mixtureto the grinding chamber through the loading port at the top of the mill.The milling was done at 1000 rpm and the mill discharged by invertingthe mill and emptying through the loading port. The recovered materialwas sieved to separate the grinding media from the powder.

Variations to the milling conditions set out above are indicated in thevariations column in the data tables. The key to these variations isshown in Table A.

Particle Size Measurement:

The particle size distribution (PSD) was determined using a MalvernMastersizer 2000 fitted with a Malvern Hydro 2000S pump unit.Measurement settings used: Measurement Time: 12 seconds, Measurementcycles: 3. Final result generated by averaging the 3 measurements.Samples were prepared by adding 200 mg of milled material to 5.0 mL of1% PVP in 10 mM hydrochloric acid (HCl), vortexing for 1 min and thensonicating. From this suspension enough was added into the dispersant(10 mM HCl) to attain a desired obscuration level. If necessary an extra1-2 minutes of sonication was applied using the internal sonicationprobe in the measurement cell. The refractive index of the activeingredient to be measured was in the range of 1.49-1.73. Any variationsto this general method are summarized in Table B.

XRD Analysis:

Powder X-Ray diffraction (XRD) patterns were measured with aDiffractometer D 5000, Kristalloflex (Siemens). The measurement rangewas from 5-18 degrees 2-Theta. The slit width was set to 2 mm and thecathode ray tube was operated at 40 kV and 35 mA. Measurements wererecorded at room temperature. The recorded traces were subsequentlyprocessed using Bruker EVA software to obtain the diffraction pattern.

TABLE A Variations to milling conditions. Only conditions reported inthe table have changed as compared to conditions reported above. MillingMedia Media Offload Variation Speed size Mass spped # Mill type (rpm)(inch) (kg) (rpm) A 1HD 1 L 0.25 B 1S 0.5 gal 5 C 1S 0.5 gal 4 D 1S 0.5gal 500 E 1S 0.5 gal 550-555 F 1S 1.5 gal 316-318 21 G 1S 1.5 gal 500 21H 1S 1.5 gal 355 21 I 1S 1.5 gal 355 18 J 1S 1.5 gal 21 K 1S 1.5 gal18.4 L 1S 1.5 gal 400 M 1S 1.5 gal 21 57 N 1S 1.5 gal 57 O 1S 0.5 gal400 400 P 1S 0.5 gal 500 350 Q HICOM ⅛ R HICOM 11.7

TABLE B Variations to particle size measurement conditions. VariationSample Measurement Addition # Dispersant Dispersant Method 1 0.1% PVP inDI water Powder addition 2 0.2% Pluronic L81 DI water in DI water 3Saturated glyphosate Powder in DI water addition 4 Saturated glyphosatePowder in DI water addition 5 1% PVP in DI water DI water 6 DI waterPowder addition 7 1% PVP in DI water Saturated creatine in DI water 8 1%PVP in DI water 10 mM HCl 9 0.2% Pluronic L81 Acidified with in DI water1M HCl 10 1% PVP in DI water 0.1% PVP in DI water 11 1% PVP in DI water1% PVP in DI water 12 Filtered before PSD measurement

Abbreviations:

HCl: Hydrochloric acid

Nap: Naproxen acid

PSD: Particles size distribution

PVP: Polyvinyl pyrrolidone

RI: Refractive index

Rpm: Revolutions per minute

SLS: Sodium lauryl sulphate

SSB: Stainless Steel Balls

XRD: X-Ray Diffraction

Other abbreviations used in the data tables are listed below in Table C(for actives), Table D (for matrices) and Table E (for surfactants). Inthe data tables single letter with example number abbreviations havebeen used to identify specific sample numbers within the table. The datatables shown in the figures the use of surfactant, matrix areinterchangeable and do not necessarily define the nature of thatmaterial.

TABLE C Abbreviations used for active pharmaceutical ingredients. APIName Abbreviation 2,4-Dichlorophenoxyacetic acid 2,4D Anthraquinone ANTCelecoxib CEL Cilostazol CIL Ciprofloxacin CIP Creatine Monohydrate CRMCyclosporin A CYA Diclofenac Acid DIC Glyphosate GLY Halusulfuron HALIndomethacin IND Mancozeb MAN Meloxicam MEL Naproxen MTX Metsulfuron METNaproxen Acid NAA Naproxen Sodium NAS Progesterone PRO Salbutamol SALSulfur SUL Tribenuran TRI

TABLE D Abbreviations used for excipients. Matrix Name AbbreviationCalcium Carbonate CAC Glucose GLU Lactose Anhydrous LAA LactoseMonohydrate LAC Lactose Monohydrate Food Grade LFG Malic Acid MAAMaltitol MAL Mannitol MAN Sodium Bicarbonate SB Sodium Chloride SCSorbitol SOR Sucrose SUC Tartaric Acid TA TriSodium Citrate DihydrateTCD Whey Powder WP Xylitol XYL

TABLE E Abbreviations used for surfactants Surfactant Name AbbreviationAerosil R972 Silica AS Benzalkonium Chloride BC Brij700 B700 Brij76 B76Cremophor EL CEL Cremophor RH-40 C40 Dehscofix 920 D920 Docusate SodiumDS Kollidon 25 K25 Kraftsperse 1251 K1251 Lecithin LEC Poloxamer 188P188 Microcrystalline Cellulose MCC Poloxamer 407 P407 PolyethyleneGlycol 3000 P3000 Polyethylene Glycol 8000 P8000 Polyoxyethylene 40Stearate P40S Polyvinyl Pyrrolidone (Kollidon 30) PVP Primellose PMLPrimojel PRI Sodium Deoxycholate SDC Sodium Dodecyl Sulphate SDS SodiumDodecylbenzenesulphonic Acid SDA Sodium N-Lauroyl Sarcosine SNS SodiumOctadecyl Sulphate SOS Sodium Pentane Sulphonate SPS Soluplus HS15 SOLTeric 305 T305 Tersperse 2700 T2700 Terwet 1221 T1221 Terwet 3785 T3785Tween 80 T80

Example 1 Spex Milling

A range of actives, matrices and surfactants in a variety ofcombinations were milled using the Spex mill. The details of thesemillings are shown in FIGS. 1A-1G together with the particle sizedistributions of actives that were milled.

These millings demonstrate that the addition of a small amount ofsurfactant to the milling matrix delivers a smaller particle sizecompared to millings of just an active and a single matrix. Someexamples of this are samples Z and AA compared to sample Y; Sample ABcompared to sample AC; sample AE compared to sample AD; sample AGcompared to sample AF; sample AP compared to sample AO; sample ARcompared to sample AQ, sample AT compared to sample AS; Samples AX, AYand AZ compared to sample AW; sample BC compared to sample BD; sample BIcompared to BH; samples BL-BR compared to sample BK; samples CS-DBcompared to sample DC. This last example is particularly noteworthy asthese millings were undertaken at 45% v/v. This demonstrates the broadapplicability of this invention. Some other examples of surfactantaddition being beneficial for size reduction are samples DD-DG and DI-DKcompared to sample DH; sample DM compared to sample DL. Other samplessuch as samples DY-EC compared to sample DX; sample AV compared tosample AU; samples B-H compared to sample A and samples K-M compared tosample J show this ti be also true when particle size statistics suchthe %<1 micron as used.

Note that this applies to mechanochemcial matrix milling as well. Thisis demonstrated by sample BI where naproxen sodium is milled withtartaric acid and converted to naproxen acid. FIG. 1H shows XRD datathat demonstrates the transformation.

Other samples such as CB-CR show examples were surfactants suitable foruse with IV formulations can be used to manufacture very smallparticles.

It is also noteworthy that samples DS and DT could be sized using asaturated solution of the active (salbutamol) demonstrating that activeswith high water solubility can be measured as long as care is taken whenmeasuring the size.

Two sets of data, samples N-Q and samples R-U, also demonstrate that theinvention described herein is unique. In these samples the active milledwith a matrix and surfactant produces small particles. When milled withmatrix alone the particles sizes are larger, in the case of sample Qthey are not even nanoparticles. When the active is milled with just 1%surfactant the resultant particle size is very large. Even when 80%surfactant is used the size is large.

Example 2 110 mL Attritor

A range of actives, matrices and surfactants in a variety ofcombinations were milled using the 110 ml stirred attritor mill. Thedetails of these millings are shown in FIG. 2A together with theparticle size distributions of actives that were milled.

These millings also demonstrate that the addition of a small amount ofsurfactant to the milling matrix delivers a smaller particle sizecompared to millings of just an active and a single matrix in a smallscale stirred mill as well as the vibratory Spex mill. Sample F alsodemonstrates that small particles can be achieved at high % actives whena surfactant is present. Sample D and E also show that the addition ofthe surfactant also increased the yield of powder from the mill.

Example 3 Second Matrix

In this example naproxen was milled with a mixture of two matrices usingthe Spex mill. The details of these millings are shown in FIG. 3Atogether with the particle size distributions of actives that weremilled. Samples A and B were milled in a primary matrix of lactosemonohydrate and 20% of second matrix. The particle size of thesemillings is smaller than the same milling with just lactose monohydrate(See example 1 sample No AH, FIG. 1B). The particle size is also smallerthan naproxen milled in the secondary matrices (See example 1 sample NoAl and AJ, FIG. 1B). This shows the mixed matrices have synergytogether.

Samples C-E were milled in anhydrous lactose with 20% of a secondmatrix. All these samples had a particle size much smaller than naproxenmilled in anhydrous lactose alone (See example 1 sample No AK, FIG. 1B).

These millings demonstrate that the addition of a second matrix to theprimary milling matrix delivers a smaller particle size compared tomillings with just a single matrix.

Example 4 1 L Attritor

Two actives with various combinations of lactose monohydrate and SDSwere milled using the 1 L stirred attritor mill. The details of thesemillings are shown in FIG. 4A together with the particle sizedistributions of actives that were milled.

Sample A and B are millings of meloxicam at 20%. While sample B has aslightly smaller particle size than sample A there is a dramaticdifference in the amount of material recovered from the milling. SampleA, milled with 3% SDS has a high yield of 90% whereas sample B with nosurfactant has practically no yield with all the powder caked in themill.

In samples C-F the milling of 13% indomethacin shows that the use of asecond matrix (tartaric acid) in combination with 1% SDS delivers thebest outcome of a good particle size and high yield. Sample D which hasjust the mixed matrix has very good particle size but poor yield.

These results show that the addition of a small amount of surfactantimproves milling performance.

Example 5 750 mL Attritor

Two actives with various combinations surfactants were milled using the750 ml stirred attritor mill. The details of these millings are shown inFIG. 5A together with the particle size distributions of actives thatwere milled.

In samples A-C three millings of naproxen are shown. Sample A has just1% SDS as a surfactant. Samples B and C have a second surfactant presentand these samples have a smaller particle size as measured by the %<500nm, %<1000 nm and %<2000 nm.

In samples D-F three millings of indomethacin are shown. Sample D hasjust 1% SDS as a surfactant. Samples E and F have a second surfactantpresent and these samples have a smaller particle size compared tosample D.

These examples demonstrate that the use of combination of surfactantscan be useful to achieve better reduction in particle size.

Example 6 ½ Gallon 1S

A range of actives, matrices and surfactants in a variety ofcombinations were milled using the ½ gallon 1S mill. The details ofthese millings are shown in FIGS. 6A-C together with the particle sizedistributions of actives that were milled.

The following examples demonstrate the increased yield obtained whenmilling an active in a ½ gallon 1S attritor mill with a surfactant ascompared to no surfactant, with all other factors being identical.Sample C and D (FIG. 6A) shows Naproxen acid milled in Mannitol withyields of 92% and 23%, with and without surfactant. Sample S and AL(FIGS. 6B and C) show the same for glyphosate with yields of 95% and26%, respectively. Sample Al and AJ (FIG. 6B) show Ciprofloxacin yieldsof 94% and 37% with and without surfactant while sample AM an AN (FIG.6C) show Celecoxib yields of 86% and 57% with and without surfactants.Finally, samples AP and AQ (FIG. 6C) shows milling Mancozeb with orwithout surfactants results in yields of 90% and 56%, respectively.

The following examples illustrates that milling an active in a ½ gallon1S attritor mill with a surfactant as compared to without surfactant andall other factors identical, leads to smaller particle size aftermilling. Sample C and D (FIG. 6A) shows a D(0.5) of 0.181 and 0.319 withor without surfactant, while sample AM and AN (FIG. 6C) shows D(0.5) of0.205 and 4.775 with and without surfactants.

The series of samples Q-S are timepoints taken from a single glyphosatemilling. The data demonstrates that the size of the actives decreaseswith milling time.

Other samples such as V-AA show examples were surfactants suitable foruse with IV formulations can be used to manufacture very smallparticles.

Some of the particle size data in FIGS. 6A-C was converted to a numberaverage particle size and is shown in the tables. This number wascalculated in the following way. The Volume distribution was transformedto the number distribution using the Malvern Mastersizer software. Foreach size bin the size of the bin was multiplied by the % of particlesin the bin. This numbers were added together and divided by 100 to givethe number average particle size.

Example 7 Naproxen

Naproxen was milled with various combinations of matrices andsurfactants using a variety of mills. The details of these millings areshown in FIG. 7A together with the particle size distributions ofactives that were milled. Samples A, B, E, G, H and I were milled in aSpex mill. Samples C, D and F were milled in the 750 ml atrittor. Theremaining samples were milled in the ½ gallon 1S mill.

Samples A compared to sample B and sample H compared to sample Gdemonstrate that the addition of one or more surfactants enables theproduction of smaller active particles. Other millings such as samplesC-F show that naproxen can be milled small at very high active loadings.Sample I shows that disintegrant can be added during milling and noteffect the production of small active particles. Note that the particlesize in sample I is after filtration through a 10 micron filter. SampleN shows an alternative way to manufacture a formulation with smallparticles and disintegrants. In this example the powder from sample Mwas left in the mill and a wetting agent (PVP) and disintegrant wereadded. The powder was milled for a further 2 minutes and then unloadedwith a very high yield of 97%.

The series of samples J-M are timepoints taken from a single milling.The data demonstrates that the size of the actives decreases withmilling time.

Example 8 Hicom

A range of actives, matrices and surfactants in a variety ofcombinations were milled using the Hicom mill. The details of thesemillings are shown in FIG. 8A together with the particle sizedistributions of actives that were milled.

The data shows that the invention described herein can be used with theHicom mill with its nutating action. The data in FIG. 8A shows that avariety of actives can be milled small in very short times and give verygood yields at 500 gram scale.

Sample N and O show that cocoa powder can be reduced to very fine sizesin short times using the invention describes here in combination withthe Hicom nutating mill. Likewise Sample P shows that this is also thecase for cocoa nibs.

Example 9 1.5 Gallon 1S

A range of actives, matrices and surfactants in a variety ofcombinations were milled using the 1.5 Gallon 1S mill. The details ofthese millings are shown in FIGS. 9A-B together with the particle sizedistributions of actives that were milled.

The following examples demonstrate the increased yield obtained whenmilling an active in a 1.5 gallon 1S attritor mill with a surfactant ascompared to no surfactant, with all other factors being identical.Sample J and N (FIG. 9A) shows yields of 51% and 80%, without and withsurfactant. Sample K and P (FIG. 9A) show yields of 27% and 80%, withoutand with surfactant, while sample L (FIG. 9A) show a yield of 94% withsurfactant and the control without surfactant (sample M, FIG. 9A)resulted in no yield due to caking within the mill.

The following examples illustrates that milling an active in a 1.5gallon 1S attritor mill with a surfactant as compared to withoutsurfactant and all other factors identical, leads to smaller particlesize after milling. Sample F and G (FIG. 9A) shows a D(0.5) of 0.137 and4.94 with or without surfactant, while sample K and P (FIG. 9A) showsD(0.5) of 0.242 and 0.152 without and with surfactants.

The series of samples Al-AL are timepoints taken from a single meloxicammilling. The data demonstrates that the size of the actives decreaseswith milling time.

Other samples such as A-E show examples were surfactants suitable foruse with IV formulations can be used to manufacture very smallparticles.

Sample M was a milling of meloxicam in lactose monohydrate withoutsurfactant. 3 minutes into the milling the mill refused to turn. Themilling was stopped and started again but only ran for another 3 minutesbefore stopping again. At this point the mill was taken apart and noevidence of caking was found. However the powder had a gritty feeling toit and was locking the medium and shaft such that it was not possible toturn. The media was weighed and it as found that 150 grams of powder wason the media indicating that it was sticking to the media and making ithard to move. At this point the mill was re-assembled and the powder andmedia put back in. 30.4 grams of SDS was included in the milling makingit similar to milling L. After the addition of the surfactant the millwas run for another 14 minutes (giving a total of 20 mins) withoutincident. After offloading the powder the media was weighed and theweigh of powder on the media was only 40.5 grams. This indicates theaddition of surfactant has improved the milling performance and abilityto mill the powder.

Some of the particle size data in FIGS. 9A-B was converted to a numberaverage particle size and is shown in the tables. This number wascalculated in the following way. The Volume distribution was transformedto the number distribution using the Malvern Mastersizer software. Foreach size bin the size of the bin was multiplied by the % of particlesin the bin. This numbers were added together and divided by 100 to givethe number average particle size.

Example 10 Large Scale 25/11 Kg

Sample A (FIG. 10A) was milled in the Siebtechnik mill for 15 minutes.After this time the powder was completely caked onto the walls of themill and the media. No powder could be removed to measure the particlesize. At this point 0.25 g (1 w/w %) SLS was added to mill chamber andmilling was then undertaken for a further 15 minutes. After the secondperiod of milling in the presence of SLS powder was no longer caked ontothe media and some free powder was also present. The observations madebefore and after the addition of the SLS demonstrate that the additionof the surfactant lessens the problem of caking. With the addition ofsurfactant the caked material could be recovered to become free powderagain with small particle size.

Sample B-E was milled in horizontal Simoloyer mills. The details ofthese millings are shown in FIG. 10A together with the particle sizedistributions of actives that were milled.

The data shows that the invention described herein can be used withSimoloyer mills with their horizontal attritor action. Of particularnote is example E which was milled at 11 kg scale. This demonstrates theinvention described herein is suitable for commercial scale milling.

Sample F was milled in a vertical attritor mill (Union Process S-30).The details of this milling is shown in FIG. 10A together with theparticle size distribution of the active milled. The data shows that theinvention described herein can be used with a S-30 mills with itsvertical attritor action. Of particular note is that this milling was at25 kg scale. This demonstrates the invention described herein issuitable for commercial scale milling.

Example 11 Naproxen

Naproxen was milled in mannitol with a range of surfactants using the ½Gallon 1S mill. The details of these millings are shown in FIG. 11Atogether with the particle size distributions of actives that weremilled.

Naproxen acid milled in Mannitol with a surfactant (Sample A, D-J inFIG. 11A) leads to higher yields, as compared to Naproxen acid milled inMannitol without surfactant (Sample K, FIG. 11A). Naproxen acid milledin Mannitol and either microcrystalline cellulose or the disintegrantprimellose (sample L or M, FIG. 11A) leads to small particle size withD(0.5) around 0.25 in both cases.

Example 12 Filtration

Some matrices, milling aids or facilitating agents that are used by thisinvention are not water soluble. Examples of these are microcrystallinecellulose and disintegrants such as croscarmellose and sodium starchglycolate. In order to more easily characterise the particle size of theactive after milling with these materials filtration methods can be usedto remove them allowing a characterisation of the active. In thefollowing examples naproxen was milled with lactose monohydrate andmicrocrystalline cellulose (MCC). The particle size was characterisedbefore and after filtration and the ability of the filters to letthrough the naproxen was demonstrated using HPLC assays. The millingdetails and the particle size are shown in FIG. 12 a. Note in this tablethe particle size with milling details is un-filtered. The particle sizein the rows with no milling details is after filtration. The sample thatwas filtered is indicated in the Active material section. The HPLCassays were performed by taking samples before and after filtrationthrough 10 micron poroplast filters. The samples taken were diluted togive a nominal concentration of 100 μg/ml. The HPLC assay data is shownin Table 12

Sample A was milled with 5% MCC. Before filtration the D50 was 2.5 μm,after filtration (sample B) the D50 was 183 nm. When sample B wasassayed the concentration was 94 μg/ml indicating that filtrationprocess retained little naproxen. A second milling (sample C) wasundertaken without MCC. The D50 was 160 nm as would be expected. Afterfiltration (sample D) the particle size was unchanged indicating that ifthe filtration process did remove any naproxen then it was removed in aneven way. Some of sample C was then milled with MCC for 1 minute. Thisis long enough to incorporate the MCC into the powder but not longenough to affect the particle size distribution. Two millings wereundertaken. Sample E incorporated 5% w/w MCC into the powder and SampleF 9% w/w. After incorporation of the MCC the particle size increaseddramatically. These samples where then filtered (Sample E and F) and thesize remeasured. After filtration the particle size is the same asSample C which was the starting material. The assay of samples E-Hindicates that filtration did not remove any naproxen of anysignificance. The combination of particle size and assay data clearlyshows that material such as MCC can easily and successfully be removedallowing the true particle size of the active to be measured.

Samples I and J were millings conducted with 10 and 20% w/w MCC. Theparticle size post filtration is show as sample K and L. Again thefiltration has delivered a reduction in particle size due to the removalof the MCC component. And again the HPLC assay of sample I-L showslittle naproxen was lost during filtration.

This data also demonstrates that MCC can successfully be used as comatrix in the invention disclosed herein.

TABLE 12 The HPLC assay of naproxen before and after filtration ofsamples. Sample No. HPLC Assay (μg/ml) B 94 D 93 E 99 F 96 G 98 H 97 I94 J 89 K 91 L 84

Example 13 Manufacture of Nanoformulation Capsules

Example 13(a) Manufacture of Naproxen (200 mg) Nanoformulation Capsules.

Nine sublots of naproxen nanoformulation milled powder were combined(Example 9, Sample Z-AH), roller compacted, processed in a Quadro®Comil®, and encapsulated. For each milling sublot, 334 g of naproxen,599 g of mannitol, 9.55 g of povidone K30, and 9.55 g of sodium laurylsulfate were charged into an 8-qt V blender and mixed for 10 minutes,yielding a powder of approximate composition 35% naproxen, 63% mannitol,1% povidone K30, and 1% sodium lauryl sulfate.

The blends were then milled individually and during the millingprocesses, unmilled material and samples were periodically dischargedand their amounts recorded. After completion of each of the individualmillings, an amount of croscarmellose sodium was added to each milling.The amount of croscarmellose sodium added was based on the theoreticalamount of milled powder remaining in the mill, such that the finalconcentration of croscarmellose sodium in the powder would be 5.38% w/wupon addition of the calculated amount. After adding the croscarmellosesodium to the attritor mill, the mill was run for 2 minutes. The milledpowder of approximate final composition 33.11% naproxen, 59.61%mannitol, 0.95% sodium lauryl sulfate, 0.95% povidone K30, and 5.38%croscarmellose sodium was then discharged from the mill.

Materials obtained from Example 9, Samples Z-AH were combined in a 1 cu.ft V-blender and mixed for 20 min. The mixed powder was processed in aFreund Model TF-156 roller compactor (screw speed=13.4, roll speed=4.1,pressure=55 kg/cm²). The powder was processed for approximately 55 min,yielding ribbons of 2.3 to 2.7 mm thickness.

The roller compacted ribbons were manually crushed and fed into thehopper of a Quadro® Comil® 197 equipped with an 1143 micron screen and0.225 inch spacer, operating at 2000 rpm. The net yield of milledgranular material was 4.183 kg.

The milled roller compacted granules were encapsulated into size 00white opaque hard gelatin capsules using a MiniCap 100 Capsule FillingMachine equipped with size 00 change parts. The capsules were filledmanually with a scraper and periodically checked for gross weight,closure integrity, and appearance. The target fill weight was 604 mg,and the average weight of an empty capsule shell was 117 mg. The filledcapsules were then polished in a capsule polishing machine. The netyield of filled, polished capsules was 4,183 g (approximately 6,925capsules).

Example 13(b): Manufacture of Indomethacin (20 Mg) NanoformulationCapsules

Indomethacin milled powder (750.0 g, Example 9, Sample T) was chargedinto the bowl of a KG-5 high shear granulator. Separately, a 30%solution of povidone K30 in purified water was prepared by dissolving47.8 g of povidone in 111.6 g of purified water.

The high shear granulator was operated with an impeller speed of 250 rpmand a chopper speed of 2500 rpm. A portion of the povidone solution(80.3 g) was introduced into the granulator over a period ofapproximately 8 minutes using a peristaltic pump. An additional 30 g ofpurified water was then added to the granulation.

After the additions of povidone solution and water were completed, thewet granules were spread on to paper-lined trays to a thickness ofapproximately ½″, and were dried in an oven at 70° C. for approximately1 hour. The granules were then manually screened through a 10 mesh handscreen, and spread on to paper-lined trays for additional drying. Thegranules were dried for a second hour, and then tested for loss ondrying; the LOD value was 1.987%.

The dried granules were processed in a Quadro CoMill (20 mesh screen,0.225 inch spacer) at 2500 rpm, yielding 689.9 g of milled granuleshaving the final composition of 12.60% indomethacin, 62.50% lactosemonohydrate, 20.86% tartaric acid, 0.95% sodium lauryl sulfate, 3.09%povidone K30.

The granules were manually filled into size 4 white opaque hard gelatincapsules using a MiniCap 100 Capsule Filling Machine set up with size 4capsule change parts. The target fill weight of each capsule was 158.7mg and the average empty capsule shell weight was 38 mg. Capsules werefilled manually using a scraper and periodically tested for grossweight. Tamping and vibration were adjusted as necessary to achieve thetarget fill weight.

The filled capsules were polished in a Capsule Polishing Machine,yielding a net weight of 803 g of filled capsules (approximately 4,056capsules).

Example 13(c): Manufacture of Indomethacin (40 Mg) NanoformulationCapsules

Two separate granulation sublots were manufactured and combined toproduce Indomethacin Nanoformulation capsules 40 mg.

Granulation sublot A was prepared by charging indomethacin milled powder(750.0 g, Example 9, Sample U) into the bowl of a KG-5 high sheargranulator. Separately, a 30% solution of povidone K30 in purified waterwas prepared by dissolving 47.8 g of povidone in 111.5 g of purifiedwater. The granulator was operated with an impeller speed of 250 rpm anda chopper speed of 2500 rpm. A portion of the povidone solution (80.3 g)was introduced into the granulator over a period of approximately 9minutes, using a peristaltic pump. An additional 20 g of purified waterwas then added to the granulation.

After the additions of povidone solution and water were completed, thewet granules were spread on to paper-lined trays to a thickness ofapproximately ½″.

Granulation sublot B was prepared by charging indomethacin milled powder(731.6 g, Example 9, Sample V and 18.4 g, Example 9, Sample U) into thebowl of a KG-5 high shear granulator. Separately, a 30% solution ofpovidone K30 in purified water was prepared by dissolving 47.8 g ofpovidone in 111.5 g of purified water. The granulator was operated withan impeller speed of 250 rpm and a chopper speed of 2500 rpm. A portionof the povidone solution (80.3 g) was introduced into the granulatorover a period of approximately 10 minutes, using a peristaltic pump. Anadditional 20 g of purified water was then added to the granulation.After the additions of povidone solution and water were completed, thewet granules were spread on to paper-lined trays to a thickness ofapproximately ½″. The wet granules from both sublots were dried in anoven at 70° C. for approximately 2.5 hours. The granules were thenmanually screened through a 10 mesh hand screen, and spread on topaper-lined trays for additional drying. The granules were dried foranother 1.5 hours, until the LOD value was 1.699%.

The dried granules were processed in a Quadro CoMill (20 mesh screen,0.225 inch spacer) at 2500 rpm. The milled granules were then added toan 8 qt V-blender and mixed for 5 minutes, yielding 1390.7 g of granuleswith a final composition of 12.60% indomethacin, 62.50% lactosemonohydrate, 20.86% tartaric acid, 0.95% sodium lauryl sulfate, 3.09%povidone K30.

An IN-CAP® automated capsule filling machine (Dott. Bonapace & C.,Milano, Italy) was set up with size (2) 16 mm dosing disc and size (2)tamping pins. Milled granules were charged into the encapsulator, alongwith size 1 white opaque hard gelatin capsule shells. The target capsulefill weight was 317.7 mg, and the average empty capsule shell weight was75 mg. Tamping pins 1-4 were all set to 9 mm, and the encapsulator wasrun at speed 2. Weight checks, closure checks, and appearance checkswere performed every 15 minutes. Filled capsules were polished in acapsule polishing machine. The net weight of filled, polished capsuleswas 1225.5 g (approximately 3,183 capsules).

Example 13(d): Manufacture of Meloxicam (7.5 Mg) NanoformulationCapsules

Milled powder (Example 9, Sample Q) was manually encapsulated using acapsule filling device (Cooper plate and capsule loader) into size “4”white-opaque hard-gelatin capsules. Upon encapsulation, each capsulecontains 7.5 mg active ingredient with a total fill weight of 105 mg.The finished capsules were packaged in 40 cc HDPE bottles (50 counts perbottle) with the bottles being enclosed using an induction seal.

Example 14 Dissolution

Example 14(a) Dissolution Rate of Milled Naproxen

The Dissolution of milled naproxen (200 mg) capsules, and commercialNaprosyn® 250 mg (naproxen) tablets (Roche Pharmaceuticals®, Inc., USA)were determined using dissolution equipment set up as USP Apparatus II(paddles) with a stirrer speed of 50 rpm. The dissolution media was 900ml of 0.3% SLS in 0.1 M sodium phosphate buffer at pH 5. The vesseltemperature was 37° C. The capsules where weighted down with a wiresinker. Six test articles were tested and the data average for each timepoint. At each time point a 1 ml sample was taken from each dissolutionvessel, filtered through a 0.45 μm filter and analyzed by HPLC. The datain Table 14a below reports the percent dissolved of the amount of activein each test article, for the specified time points.

TABLE 14a Dissolution Profiles of Naprosyn ® Tablets 250 mg and NaproxenNanoformulation Capsules 200 mg Percent of Label Claim Dissolved (%)Naproxen Naprosyn Tablets Nanoformulation Capsules Time 250 mg 200 mg 00 0 5 24 19 10 40 53 15 49 77 20 55 90 45 73 98 60 79 99

The results demonstrate that the milled naproxen capsules dissolve morequickly and more completely than the commercial reference naproxen.Those of skill in the art will readily appreciate the advantagesconferred by more rapid dissolution—more active agent is available atany given time point. Put another way, an equal quantity of dissolvednaproxen may be obtained with an initially smaller dosage amount ofmilled naproxen, as opposed to the larger initial dose required for thereference naproxen to reach to the same quantity of dissolved naproxen.Additionally, as the results make clear, the reference naproxen does notachieve complete dissolution even by the final time point, while themilled naproxen achieves greater than 90% dissolution within 20 minutes,and substantially complete dissolution by the 45 minute time point.Again, a smaller dose of milled naproxen yields a quantity of dissolvednaproxen for which a larger dose of reference naproxen would be requiredto equal.

Example 14(b): Dissolution Rate of Milled Indomethacin

In this example, dissolution rate is compared between 20 mg and 40 mgnanoformulations of the invention (Example 13(b) and 13(c)), andcommercial reference indomethacin USP 25 mg capsules (MylanPharmaceuticals Inc). The dissolution was performed using Apparatus I(baskets) according to USP <711>. The dissolution medium (900 ml at 37°C.) was 100 mM citric acid buffer (pH 5.5±0.05); The apparatus wasstirred at 100 rpm. Sampling times were 5, 10, 20, 30, 45, and 60 minplus an additional time point at 75 min (250 rpm). Sample of 8 mL weretaken and filtered through a 0.45 μm PVDF filter. The samples were assayby UV-visible spectroscopy with a detection wavelength=319 nm. The datain Table 14b below reports the percent dissolved of the amount of activein each test article, for the specified time points.

TABLE 14b Dissolution Profiles of Indomethacin Capsules USP (25 mg) andIndomethacin Nanoformulation Capsules (20 mg and 40 mg) Percent of LabelClaim Dissolved (%) Indomethacin Indomethacin Indomethacin capsulesNanoformulation Nanoformulation Time (min) USP, 25 mg Capsules 20 mgCapsules 40 mg 0 0 0 0 5 20 47 31 10 28 83 66 20 36 99 93 30 40 100 9645 43 100 96 60 46 101 97 75 63 101 97

The results demonstrate that the nanomilled indomethacin capsulesdissolve more quickly and more completely than the commercial referenceindomethacin. These same capsules were also tested in a in-vivo humanclinical trial (as described in patent application, “A novel formulationof indomethacin”, filed as PCT/AU2010/______ claiming priority to AUprovisional application 2009901740) This trial (fasted leg) demonstratedthat the 20 and 40 mg nanomilled indomethacin had faster onset comparedto the commercial reference (50 mg) (T_(max)=1.1 hours for 20 mg nano,1.25 hours for 40 mg nano and 2.0 hours for 50 mg reference) and that 40mg nanomilled indomethacin had higher a higher Cmax compared to thecommercial reference (50 mg) (C_(max)=2995 ng/ml for 40 mg nano and 2652ng/ml for 50 mg reference). These in-vivo data demonstrate that thein-vitro dissolution test is indicative of the behaviour of a NSAIDmanufactured using this invention.

Example 14(c): Dissolution Rate of Milled Meloxicam

In this example, dissolution rate is compared between a 7.5 mgnanoformulation of this invention (Example 13(d)), and two commercialreference products Mobicox® 7.5 mg Tablets and Mobic® 7.5 mg Capsules(Both Boehringer Ingelheim). Dissolution was performed using ApparatusII (paddles) according to USP <711>. The dissolution medium was 10 mMphosphate buffer (pH 6.1) with 0.1% w/w sodium lauryl sulfate (500 ml at37° C.). The apparatus was stirred at 50 rpm. Samples were taken atvarious time points from 5 to 60 minutes. For each sample 1 mL wastaken, filtered through a 0.45 μm filter and assayed by HPLC using adetection wavelength of 362 nm. The data in Table 14c below report thepercent dissolved of the amount of active in each test article, for thespecified time points.

TABLE 14C Dissolution profiles of Commercial Meloxicam Tablets andCapsules and Meloxicam Nanoformulation Capsules Percent of Label ClaimDissolved (%) Meloxicam Mobicox ® Mobic ® Capsules Nanoformulation Time(min) Tablets 7.5 mg 7.5 mg Capsules 7.5 mg 0 0 0 0 5 39 19 44 10 50 4368 15 57 52 20 82 30 66 64 86 45 89 60 73 72 93

The results demonstrate that the milled meloxicam capsules dissolve morequickly and more completely than the commercial reference meloxicam. Thecapsules tested in this dissolution study were also tested in a in-vivohuman clinical trial (as described in patent application, “A novelformulation of meloxicam”, PCT/AU2010/______, claiming priority to AUprovisional application 2009901742). This trial (fasted leg)demonstrated that the 7.5 mg nanomilled meloxicam had faster onsetcompared to the commercial reference (Tmax=2.0 hours for nano, 5.0 hoursreference) and that nanomilled meloxicam had higher a higher Cmaxcompared to the commercial reference (Cmax=1087 ng/ml for nano and 628ng/ml for reference). These in-vivo data demonstrate that the in-vitrodissolution test is indicative of the behaviour of a NSAID manufacturedusing this invention.

Example 15 Bioavailability of Milled Naproxen

This Example describes a Single-Dose, Four-Way Crossover, RelativeBioavailability Study of Naproxen Nanoformulation Capsules (200 mg) inHealthy Subjects under Fed and Fasted Condition.

The pharmacokinetic study described in this example uses NaproxenNanoformulation Capsules manufactured as described in Example 13.

Naprosyn® (naproxen) is a nonsteroidal anti-inflammatory drug (NSAID)with analgesic and antipyretic properties. The mechanism of action ofthe naproxen anion, like that of other NSAIDs, is not completelyunderstood but may be related to prostaglandin synthetase inhibition.

Naproxen is rapidly and completely absorbed from the gastrointestinaltract with an in vivo bioavailability of 95%. The elimination half-lifeof naproxen ranges from 12 to 17 hours.

After administration of Naprosyn® tablets, peak plasma levels areattained in 2 to 4 hours. Naproxen has a volume of distribution of 0.16L/kg. At therapeutic levels naproxen is greater than 99% albumin-bound.

Naproxen is extensively metabolized to 6-O-desmethylnaproxen, and bothparent and metabolites do not induce metabolizing enzymes. Both naproxenand 6-O-desmethylnaproxen are further metabolized to their respectiveacyl glucuronide conjugated metabolites.

The clearance of naproxen is 0.13 mL/min/kg. Approximately 95% of thenaproxen from any dose is excreted in the urine, primarily as naproxen(<1%), 6-O-desmethyl naproxen (<1%) or their conjugates (66% to 92%).The plasma half-life of the naproxen anion in humans ranges from 12 to17 hours. The corresponding half-lives of both naproxen's metabolitesand conjugates are shorter than 12 hours, and their rates of excretionhave been found to coincide closely with the rate of naproxendisappearance from the plasma. Small amounts, 3% or less of theadministered dose, are excreted in the feces.

In patients taking naproxen in clinical trials, the most frequentlyreported adverse experiences in approximately 1% to 10% of patients are:gastrointestinal (GI) experiences, including: heartburn, abdominal pain,nausea, constipation, diarrhea, dyspepsia, stomatitis; central nervoussystem: headache, dizziness, drowsiness, lightheadedness, vertigo;dermatologic: pruritus (itching), skin eruptions, ecchymoses, sweating,purpura; special senses: tinnitus, visual disturbances, hearingdisturbances; cardiovascular: edema, palpitations; general: dyspnea,thirst.²

Objectives

The objective of this single-dose, open-label, randomized, 5-period,5-treatment crossover study is to evaluate the relative bioavailabilityand pharmacokinetics of a test formulation of naproxen 400 mg under fedand fasting conditions, and a test formulation of naproxen 200 mg underfasting conditions, compared to a 500 mg oral dose of the commerciallyavailable reference product, Naprosyn® manufactured by RochePharmaceuticals under fed and fasting conditions.

The primary objectives of the study are:

To determine the relative bioavailability of naproxen from the 1×200 mgand 2×200 mg Test capsules versus the 500 mg Reference tablet whenadministered to healthy subjects under fasted conditions.

To determine the effect of food on the rate and extent of absorption ofa single dose of the 2×200 mg Test capsule formulation of naproxennanoformulation administered to healthy subjects.

To determine the effect of food on the rate and extent of absorption ofa single dose of the 500 mg Reference tablet formulation of naproxenadministered to healthy subjects.

To evaluate the dose proportionality between a single 200 mg Testcapsule and a 400 mg (2×200 mg capsules) dose of naproxennanoformulation administered to healthy subjects under fastingconditions.

Study Design Summary

This is a single-dose, open-label, randomized, 5-period, 5-treatmentcrossover study in which 40 healthy adult subjects will receive 5separate single-dose administrations of naproxen. Subjects receiving thefed treatments will be administered the study drug after an overnightfast of at least 10 hours, followed by consumption of an FDA standardhigh-calorie, high-fat breakfast meal beginning 30 minutes prior to eachdose.

Subjects receiving the fasting treatments will be administered the studydrug following an overnight fast of at least 10 hours.

Subjects will be assigned numbers in an ascending order, based onsuccessful completion of the screening process.

Subjects will receive each of the treatments listed below in randomizedfashion during the five treatment periods:

Treatment A: Test Formulation Fed Naproxen conditions Dose = 2 × 200 mgcapsule Treatment B: Test Formulation Fasting Naproxen conditions Dose =2 × 200 mg capsule Treatment C Test Formulation Fasting Naproxenconditions Dose = 1 × 200 mg capsule Treatment D Reference Product FedNaprosyn ® conditions Dose = 1 × 500 mg tablet Roche PharmaceuticalsTreatment E Reference Product Fasting Naprosyn ® conditions Dose = 1 ×500 mg tablet Roche Pharmaceuticals

Each drug administration will be separated by a washout period of atleast 7 days. Treatments A and D will be orally administered along with240 mL (8 fl. oz.) of room temperature tap water following a 10-hourovernight fast and standard high-fat, high-calorie breakfastadministration. Treatments B, C, and E will be orally administered alongwith 240 mL (8 fl. oz.) of room temperature tap water following a10-hour overnight fast.

After dosing, no food will be allowed until 4 hours post-dose. Exceptfor the 240 mL of room temperature tap water provided with the dose, nowater may be consumed for 1 hour prior through 1 hour post dose. Waterconsumption will follow the guidelines in Section 5.4. With theexception of the standard high-fat, high-calorie breakfast meal servedwith Treatments A and D, meals will be the same and scheduled atapproximately the same times relative to dose for each study period.

Subjects who withdraw from the study will not be replaced.

During each study period, 6 mL blood samples will be obtained prior toeach dosing and following each dose at selected times through 72 hourspost-dose. A total of 115 pharmacokinetic (PK) blood samples will becollected from each subject, 23 samples in each study period. Plasmapharmacokinetic samples will be analyzed for naproxen using a validatedanalytical method. Appropriate pharmacokinetic parameters will becalculated for each formulation using non-compartmental methods. Inaddition, blood will be drawn and urine will be collected for clinicallaboratory testing at screening and at the end of the study.

Subject Selection

-   -   Inclusion Criteria    -   All subjects must satisfy the following criteria to be        considered for study participation:        -   Subject must be a male or non-pregnant, non-breastfeeding            female.        -   Subject must be between 18 and 55 years of age (inclusive).        -   Subject's Body Mass Index (BMI) must be between 18 and 30            kg/m² (inclusive), and subject must weigh a minimum of 50 kg            (110 lbs).        -   Female subjects must agree to use one of the following forms            of birth control from screening until 14 days after            completion of the study:            -   Vasectomized partner (at least 6 months prior to dosing)            -   Post-menopausal (at least 2 years prior to dosing)            -   Surgically sterile (bilateral tubal ligation,                hysterectomy, bilateral oophorectomy) at least 6 months                prior to dosing            -   Double barrier (diaphragm with spermicide; condoms with                spermicide)            -   IUD (intra-uterine device)            -   Abstinence (must agree to use a double barrier method if                they become sexually active during the study)            -   Implanted or intrauterine hormonal contraceptives in use                for at least 6 consecutive months prior to study dosing                and throughout the study duration            -   Oral, patch, and injected contraceptives in use for at                least 3 consecutive months prior to study dosing and                throughout the study duration.        -   Subject must voluntarily consent to participate in this            study and provide their written informed consent prior to            start of any study-specific procedures.        -   Subject is willing and able to remain in the study unit for            the entire duration of each confinement period and return            for outpatient visits.        -   Subject is willing and able to consume the entire            high-calorie, high-fat breakfast meal in the designated            timeframe required when assigned to a fed study period study            period.    -   Exclusion Criteria    -   Subjects will be excluded for any of the following:    -   History or presence of clinically significant cardiovascular,        pulmonary, hepatic, renal, hematologic, gastrointestinal,        endocrine, immunologic, dermatologic, neurologic, oncologic, or        psychiatric disease or any other condition that, in the opinion        of the Investigator, would jeopardize the safety of the subject        or the validity of the study results.    -   Specifically, subjects with history or presence of congestive        heart failure, coronary artery disease, fluid retention,        hypertension, ulcer disease or gastrointestinal bleeding, active        kidney disease, or bleeding disorder.    -   Has a clinically significant abnormal finding on the physical        exam, medical history, ECG, or clinical laboratory results at        screening.    -   History or presence of allergic or adverse response to naproxen        or related drugs.    -   Has been on a significantly abnormal diet during the 4 weeks        preceding the first dose of study medication.    -   Has donated blood or plasma within 30 days prior to the first        dose of study medication.    -   Has participated in another clinical trial within 30 days prior        to the first dose of study medication.    -   Has used any over-the-counter (OTC) medication, including        nutritional supplements, within 7 days prior to the first dose        of study medication.    -   Has used any prescription medication, except hormonal        contraceptive or hormonal replacement therapy, within 14 days        prior to the first dose of study medication.        -   Subjects that have discontinued the use of implanted,            intrauterine, or injected hormonal contraceptives must not            have used any for 6 months prior to study start.        -   Subjects that have discontinued the use of oral or patch            hormonal contraceptives must not have used any for 1 month            prior to study start.        -   Has been treated with any known enzyme altering drugs, such            as barbiturates, phenothiazines, cimetidine, carbamazepine,            etc., within 30 days prior to the first dose of study            medication.        -   Has smoked or used tobacco products within 60 days prior to            the first dose of study medication.        -   Has any prior history of substance abuse or treatment            (including alcohol) within the past 2 years.        -   Is a female with a positive pregnancy test result.        -   Has a positive urine screen for drugs of abuse            (amphetamines, barbiturates, benzodiazepines, cocaine,            cannabinoids, opiates).        -   Has had a positive test for, or has been treated for            hepatitis B, hepatitis C or HIV.    -   Restrictions        -   Subject must not take any OTC medication, including            nutritional supplements, within 7 days prior to the first            dose of study medication until the end-of-study visit            without evaluation and approval by the study investigator.        -   Subject must not take any prescription medication, with the            exception of female hormonal contraceptives or hormone            replacement therapy, from 14 days prior to the first dose of            study medication until the end-of-study visit without            evaluation and approval by the study investigator.        -   Subject must not consume beverages and foods containing            alcohol, grapefruit, or caffeine/xanthine from 48 hours            prior to the first dose of study medication until the            end-of-study visit. Subjects will be instructed not to            consume any of the above products; however, allowance for an            isolated single incidental consumption may be evaluated and            approved by the study investigator based on the potential            for interaction with the study drug.        -   Subject must not donate blood or plasma 30 days prior to the            first dose of study medication until the end-of-study visit.            It is recommended that blood/plasma donations not be made            for at least 30 days after the end-of-study visit.        -   Subject must not use tobacco products from 60 days prior to            the first dose of study medication until the end-of-study            visit.        -   Subject must not engage in strenuous exercise from 48 hours            prior to the first dose of study medication until the            end-of-study visit.        -   Female subjects must utilize one of the following forms of            contraception, if sexually active with a male partner, from            screening until 14 days after completion of the study.            Approved forms of contraception are:            -   Vasectomized partner (at least 6 months prior to dosing)            -   Post-menopausal (at least 2 years prior to dosing)            -   Surgically sterile (bilateral tubal ligation,                hysterectomy, bilateral oophorectomy) at least 6 months                prior to dosing            -   Double barrier (diaphragm with spermicide; condoms with                spermicide)            -   IUD (intra-uterine device)            -   Abstinence (must agree to use a double barrier method if                they become sexually active during the study.)            -   Implanted or intrauterine hormonal contraceptives must                be used for at least 6 consecutive months prior to study                dosing and throughout the study duration            -   Oral, patch, and injected contraceptives must be used                for at least 3 consecutive months prior to study dosing                and throughout the study duration.        -   Subjects who have discontinued the use of implanted,            intrauterine, or injected hormonal contraceptives must not            have used any for 6 months prior to study start.        -   Subjects who have discontinued the use of oral or patch            hormonal contraceptives must not have used any for 1 month            prior to study start.    -   Screening    -   Each potential study participant will have the following        assessments by the Investigator or designee within 28 days prior        to study start: medical history and demographic data, including        sex, age, race, ethnicity, body weight (kg), height (cm), BMI        (kg/m²), and smoking habits. Each potential participant will        receive a physical examination, electrocardiogram (ECG), and the        laboratory tests for hematologic, hepatic, and renal function        listed below. ECGs will be performed after subject has been in        supine position for a minimum of 5 minutes. All potential        subjects will be tested for hepatitis B, hepatitis C, and Human        Immunodeficiency Virus (HIV) at screening. Urine drug screen        tests will be conducted on all potential subjects. Serum        pregnancy tests will be conducted on all female subjects.    -   Only medically healthy subjects with clinically acceptable        laboratory profiles and ECGs will be enrolled in the study. The        informed consent documents will be discussed with each potential        participant, and each individual will sign an informed consent        document for the study prior to any study-specific procedures        being performed.    -   A positive test result for pregnancy, HIV, hepatitis B,        hepatitis C, or urine drug screen will end the screening        process.    -   Laboratory Tests    -   A Clinical Laboratory Improvement Amendments (CLIA) certified        laboratory will perform the following clinical laboratory tests        for this study:        -   Hematology        -   The following will be evaluated: hemoglobin, hematocrit,            total and differential leukocyte count, red blood cell count            (RBC), and platelet count.        -   Serum Chemistry        -   The following will be evaluated: albumin, blood urea            nitrogen (BUN), creatinine, total bilirubin, alkaline            phosphatase (ALP), aspartate transaminase (AST), alanine            transaminase (ALT), sodium (Na⁺), potassium (K⁺), chloride            (Cl⁻), lactate dehydrogenase (LDH), calcium (Ca), uric acid,            and glucose.        -   Serology        -   Blood will be tested for Hepatitis B Surface Antigen,            Hepatitis C Antibody, and Human Immunodeficiency Virus            (HIV).        -   Urinalysis        -   The following will be evaluated by an automated or manual            urine “dipstick” method: pH, specific gravity, protein,            glucose, ketones, bilirubin, blood, nitrite, leukocyte            esterase, and urobilinogen. If protein, occult blood,            nitrite, or leukocyte esterase values are out of range, a            microscopic examination will be performed.            -   Urine Drug and Alcohol Screens        -   Urine samples will be tested for drugs of abuse            (amphetamines, benzodiazepines, barbiturates, cannabinoids,            cocaine, opiates) at screening. Urine samples will be tested            for drugs of abuse and alcohol at each check-in.        -   Pregnancy Test (Female Subject Only)        -   A serum pregnancy test will be performed on all female            subjects at screening. A urine pregnancy test will be            performed on all female subjects at each check-in.

Study Procedures

-   -   Subject Assignment    -   Forty subjects will be dosed in this study. Each subject will        receive an assigned treatment sequence based on the        randomization schedule prepared by the clinical site. Subjects        will be randomized to receive either Treatment A, B, C, D, or E        during the first study period. After a minimum washout of 7        days, each subject will crossover to receive an alternate        treatment. At the completion of the study, each subject will        have received a single dose of Treatment A, a single dose of        Treatment B, a single dose of Treatment C, a single dose of        Treatment D, and a single dose of Treatment E.

Sequence Period 1 Period 2 Period 3 Period 4 Period 5 Number TreatmentTreatment Treatment Treatment Treatment 1 A B C D E 2 B C D E A 3 C D EA B 4 D E A B C 5 E A B C D

-   -   The maximum duration of the study from screening to study exit        will be approximately 59 days.    -   Check-In Procedures    -   All subjects will be asked to affirm that the exclusion criteria        and restrictions have not been violated since the screening. The        subjects' responses will be documented.    -   A urine sample will be collected from all subjects at each study        check-in to screen for drugs of abuse (UDS) and alcohol. If at        any time the drug or alcohol test is positive, the subject will        be discontinued from study participation.    -   A urine sample will be collected from all female subjects for a        urine pregnancy test at each check-in. This test must be        negative for the subject to continue study participation.    -   Confinement    -   Subjects will be admitted to the research center at an        appropriate time the evening prior to study drug administration        to ensure a minimum 10-hour fast. Subjects will remain in the        research center until completion of the 24-hour procedures for        each study period and return for outpatient visits at        approximately 36, 48, and 72 hours post-dose in each study        period.    -   Fasting/Meals/Beverages        -   Fed Treatments (A and D)        -   An optional snack will be served the evening of check-in.            All subjects will then be required to fast for at least 10            hours prior to consuming a standard breakfast. Subjects will            receive a required FDA standard high-fat, high-calorie            breakfast to begin 30 minutes prior to scheduled            administration of the dose and to end (last bite taken)            within 5 minutes prior to dosing. The subjects will fast for            4 hours thereafter. Standard meals will be provided at            approximately 4 and 10 hours after drug administration and            at appropriate times thereafter. Meal/snack menus will be            the same for all study periods.        -   The following high-fat (approximately 50% of total caloric            content of the meal), high-calorie (approximately 1000            calories) breakfast will be ingested approximately 30            minutes prior to administration of the drug.        -   2 eggs fried in butter        -   2 strips of bacon        -   2 slices of toast with butter        -   4 ounces of hash brown potatoes        -   8 ounces of whole milk        -   This meal contains approximately 150 protein calories, 250            carbohydrate calories, and 500-600 fat calories. An            equivalent meal may be substituted with documentation of the            menu and caloric contents.        -   Water will be allowed ad lib during the study except for 1            hour prior through 1 hour post dose.        -   Fasting Treatments (B, C, and E)        -   An optional snack will be served the evening of check-in.            All subjects will then be required to fast for at least 10            hours prior to scheduled administration of the dose.            Standard meals will be provided at approximately 4 and 10            hours after drug administration and at appropriate times            thereafter. Meal/snack menus will be the same for all study            periods.        -   Water will be allowed ad lib during the study except for 1            hour prior through 1 hour post dose.    -   Drug Administration    -   Each subject will receive the oral dose of the assigned naproxen        formulation with 240 mL (8 fl. oz.) of room temperature tap        water. Subjects must swallow the study medication intact. The        medication should NOT be crushed or chewed. A mouth check will        be performed immediately after dose to ensure that the        medication has been appropriately swallowed.    -   The subjects will remain seated, except as otherwise required        for study procedures or personal needs, for the first 4 hours        after dosing. Subjects will not be allowed to lie down, except        as directed by clinical staff secondary to adverse events, for        the first 4 hours after dosing.    -   Blood Sampling, Processing and Shipment    -   A total of 690 mL (115×6 mL samples) will be collected for PK        analysis. In addition, approximately 40 mL of blood will be        collected for screening and the end-of-study clinical laboratory        evaluations. The total volume of blood collected will not exceed        730 mL.    -   Blood samples (1×6 mL) will be collected in vacutainer tubes        containing K₂EDTA as a preservative, at 0 (pre-dose) and at        0.25, 0.5, 0.75, 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.5,        4, 5, 8, 12, 16, 24, 36, 48, and 72 hours after dosing. The        pre-dose blood sample will be collected within 60 minutes prior        to each dose of study drug. Pre-dose blood samples obtained from        backup subjects who are randomized into the study may exceed the        pre-dose collection window. The time and date of collection for        each sample will be recorded.    -   Blood samples will be centrifuged at approximately 3000 rpm for        10 minutes at 4 degrees Centigrade. The resulting plasma samples        will be harvested and transferred into appropriately labeled        polypropylene screw-cap tubes. PK samples will be placed in a        storage freezer at minus 20 degrees Centigrade or lower within        60 minutes of blood draw. Samples will remain frozen until        assayed. A more detailed description of plasma sample        preparation requirements may be provided by the analytical        laboratory. If such a description is provided, the method of        sample preparation provided by the laboratory shall supersede        those provided in this protocol and appropriate documentation        shall be placed in the study master file.    -   The samples will be transferred to the analytical laboratory        after completion of the study or at mutually agreed upon time        points during the clinical conduct of the study. Prior to        shipment, the samples will be appropriately packed in a        Styrofoam® cooler containing dry ice. Sufficient dry ice will be        added to ensure that the samples will remain frozen for at least        24 hours for local shipments and for at least 72 hours for        remote shipments. The shipment will be accompanied by        documentation containing the following information: name of the        study drug product, protocol number, number of subjects, and        number of samples included in the shipment.    -   End-of-Study Procedures    -   Vital signs (blood pressure, pulse rate, respiration rate, and        temperature) will be measured prior to the collection of the        72-hour blood sample at Study Period 5. Following the collection        of the 72-hour blood sample at Study Period 5, all subjects will        undergo a physical examination and ECG. The ECG will be        performed after subject has been in supine position for a        minimum of 5 minutes. Blood and urine will be collected for the        same hematology, chemistry, and urinalysis tests performed        during screening. When possible, end-of-study procedures will be        performed in the event of a subject's early termination from the        study.    -   Safety Monitoring and Procedures    -   At screening, prior to each administration of naproxen, and at        the end-of-study visit (prior to last PK blood collection) the        following vital signs will be measured:        -   blood pressure        -   pulse rate        -   respiration rate        -   temperature    -   For purposes of qualifying any given subject for study        participation, out-of-range vital signs may be repeated once.    -   At approximately 2, 4, 24 and 72 hours after each dose of study        drug the following vital signs will be collected:        -   blood pressure        -   pulse rate    -   Additional vital signs measurements may be performed as deemed        medically necessary by research personnel. All vital signs        measurements will be taken after the subject has completed a        minimum 3-minute sit.    -   Subjects will be closely monitored during each confinement        period in the research facility. Subjects will remain seated,        except as otherwise required for study procedures or personal        needs, for the first four hours after dosing. Should the need to        move about occur during the first four hours after each dose,        subjects may be escorted to such procedures or activities by        research personnel as deemed medically necessary.    -   Subjects will be instructed to inform the study physician and/or        research personnel of any adverse events (AEs) that occur at any        time during the study.    -   Medical emergency personnel trained in advanced cardiac life        support will be on site to monitor subjects during the        confinement period in the research center. Emergency medical        equipment including but not limited to intubation equipment and        pulse oximetry shall be maintained on site to administer        appropriate medical care should it be required. A physician will        remain on site for a minimum of 4 hours after each dose        administration and will be available immediately by cell phone        or pager thereafter.

Adverse Events

Subjects will be monitored for any adverse events from the beginning ofconfinement until the end-of-study visit. The Investigator or amedically qualified designee will review each event and assess itsrelationship to the study drug. Each sign or symptom will be graded forseverity, and the date and time of onset, cessation and resolution willbe recorded. Treatment of any adverse reactions will be evaluated andmanaged by a physician, either at the study site or at a nearby hospitalemergency room, as appropriate.

-   -   Definitions        -   Adverse Event (AE)        -   An AE is any untoward medical occurrence in a patient or            clinical investigation subject administered a pharmaceutical            product that does not necessarily have a causal relationship            with the product. An AE can therefore be any unfavorable and            unintended sign (including a new, clinically important            abnormal laboratory finding), symptom, or disease,            temporally associated with the product, whether or not            related to the product.        -   Abnormal results of diagnostic procedures, including            laboratory findings, are considered to be AEs if the            abnormality:            -   results in study withdrawal            -   is associated with a serious adverse event (SAE)            -   is associated with clinical signs or symptoms            -   is considered by the physician to be of clinical                significance        -   The relationship to the study treatment is characterized as:

TERM DEFINITION CLARIFICATION Un- This category applies related to thoseadverse events which, after careful consideration, are clearly andincontro- vertibly due to extraneous causes (disease, environment, etc.)Pos- This category applies An adverse experience may be sibly to thoseadverse events considered possibly related if for which, after carefulor when (at least two of the medical consideration following): at thetime they are It follows a reasonable temporal evaluated, a connectionsequence from administration of with the Investigational theInvestigational Medicinal Medicinal Product (IMP) Product (IMP).administration appears It could not readily have been unlikely butcannot be produced by the subject's ruled out with certainty. clinicalstate, environmental or toxic factors, or other modes of therapyadministered to the subject. It follows a known pattern of response tothe IMP. Prob- This category applies An adverse experience may be ablyto those adverse events considered probably related if which, aftercareful or when (at least three of the medical consideration atfollowing): the time they are It follows a reasonable temporalevaluated, are felt with sequence from administration of a high degreeof certainty the IMP. to be related to the IMP. It could not bereasonably explained by the known character- istics of the subject'sclinical state, environmental or toxic factors or other modes of therapyadministered to the subject. It disappears or decreases on cessa- tionor reduction in dose. There are important exceptions when an ad- verseevent does not disappear upon discontinuation of the drug, yetdrug-relatedness clearly exists. It follows a known pattern of res-ponse to the IMP.

-   -   -   Serious Adverse Events (SAE)        -   A serious AE (SAE) is any untoward medical occurrence that            at any dose:            -   Results in death            -   Is life threatening            -   Requires inpatient hospitalization or prolongation of                existing hospitalization            -   Results in persistent or significant                disability/incapacity            -   Is a congenital anomaly            -   Is an important medical event

    -   Medical and scientific judgment should be exercised in deciding        whether it is appropriate to consider other situations serious,        such as important medical events that may not be immediately        life threatening or result in death or hospitalization but may        jeopardize the subject or may require intervention to prevent        another of the outcomes listed in the definition above.        -   Examples of such events are intensive treatment in an            emergency room or at home for allergic bronchospasm, blood            dyscrasias, or convulsions that do not result in            hospitalization, or development of drug dependency or drug            abuse.        -   An elective hospital admission to treat a condition present            before exposure to the study drug, or a hospital admission            for a diagnostic evaluation of an AE, does not qualify the            condition or event as an SAE.        -   A newly diagnosed pregnancy in a subject who has received a            study drug is not considered an SAE unless it is suspected            that the study drug interacted with a contraceptive method            and led to the pregnancy. A congenital anomaly in an infant            born to a mother who was exposed to the study drug during            pregnancy is an SAE.        -   The investigator must report all SAEs immediately, and no            later than 24 hours after first becoming aware of the event            by completing the SAE form. At the time of first            notification of an SAE, the following information should be            provided by the study site if available:            -   Subject's study number and initials            -   Subject's date of birth            -   Subject's gender            -   Date of first dose of study drug(s)            -   Date of last dose of study drug(s), if applicable            -   AE term            -   Time and date of occurrence of the event            -   A brief description of the event, outcome to date, and                any actions taken            -   The seriousness criteria(on) that were met            -   Concomitant medication at onset of the event            -   Relevant medical history information            -   Relevant laboratory test findings            -   Investigator's opinion of the relationship to study                drug. (“Is there a reasonable possibility that the study                drug caused the SAE? Yes or no?”).            -   Whether and when the subject's treatment assignment was                unblinded        -   Any missing or additional relevant information concerning            the serious (or unexpected) AE should be provided in a            written follow-up report.        -   The investigator is required to comply with applicable            regulations regarding the notification of his/her IRB or            IEC.            -   Pregnancy        -   All women of reproductive potential who participate in the            trial should be counseled on the need to practice adequate            birth control and on the importance of avoiding pregnancy            during study participation. Women should be instructed to            contact the investigator or study staff immediately if            pregnancy occurs or is suspected.            -   Follow-Up of Subjects with an Adverse Event        -   Any AE will be followed to a satisfactory resolution, until            it becomes stable, or until it can be explained by another            known cause(s) (ie, concurrent condition or medication) and            clinical judgment indicates that further evaluation is not            warranted. All findings relevant to the final outcome of an            AE must be reported in the subject's medical record.

General Considerations

-   -   Basic Principles    -   This research will be carried out in accordance with the        protocol, good clinical practices (GCPs), and applicable        regulatory requirements(s) including clinical research        guidelines established by the Basic Principles defined in the        U.S. 21 CFR Parts 50, 56, and 312 and the principles enunciated        in the Declaration of Helsinki (revised version Seoul 2008).    -   Institutional Review Board    -   This protocol will be reviewed by an appropriate IRB and study        enrollment will not commence until the Board has approved the        protocol or a modification thereof. The Board is constituted and        operates in accordance with the principles and requirements        described in the U.S. Code of Federal Regulations (21 CFR Part        56).    -   Informed Consent    -   Written informed consent will be obtained from each subject        prior to performing any baseline study-specific evaluations. The        informed consent document is prepared by the Investigator or        designee, subject to review and approval by the Sponsor, and        forwarded to a qualified IRB for final review and approval. The        IRB-approved document must contain, at minimum, the eight basic        elements of informed consent. Only the most recently        IRB-approved Informed Consent Document must be used to consent        prospective study subjects. One copy of the signed and dated        informed consent document will be given to the subject and the        original retained by the Investigator/site.

Indications for Subject Withdrawal

-   -   Subjects will be free to withdraw at any time for any reason, or        they may be withdrawn if necessary, to protect their health and        safety or the integrity of the study data. The final report will        include reasons for withdrawals.    -   Termination of the Study    -   The Principal Investigator reserves the right to terminate the        study in the interest of subject safety and welfare. The Sponsor        reserves the right to terminate the study at any time for        administrative reasons.

Documentation

-   -   All documents pertaining to the study, including a copy of the        approved protocol, copy of the informed consent document and        Health Insurance Portability and Accountability Act (HIPAA)        documents, completed case report forms (where applicable), drug        accountability and retention records, and other study related        documents will be retained in the permanent archives of the        study site. These will be available for inspection at any time        by the Sponsor or the FDA. Per 21 CFR 312, record retention for        this study is required for a period of 2 years following the        date on which this study agent is approved by the FDA for the        marketing purposes that were the subject of this investigation;        or, if no application is to be filed or if the application is        not approved for such indication, until 2 years following the        date on which the entire study (not merely the Investigator's        portion of the study, if it involved more than one investigator)        is completed, terminated, or discontinued, and the FDA is        notified.

Pharmacokinetic Analysis

-   -   Analytical Methodology    -   A full validation of a sensitive LC-MS-MS assay for naproxen in        plasma, including precision, accuracy, reproducibility, and        selectivity, will be provided to the Sponsor. The validation        report will include the stability of frozen samples, limit of        quantitation, recovery, and Watson LIMS summary tables. The        samples from all evaluable subjects completing at least one        study period will be analyzed.    -   Pharmacokinetic Analysis    -   Pharmacokinetic parameters for naproxen will be calculated using        non-compartmental analysis. The following pharmacokinetic        parameters will be determined:    -   The maximum plasma concentration (C_(max)) and time to C_(max)        (T_(max)) will be taken directly from the data. The elimination        rate constant, λ_(z), will be calculated as the negative of the        slope of the terminal log-linear segment of the plasma        concentration-time curve; the range of data to be used will be        determined by visual inspection of a semi-logarithmic plot of        concentration vs. time. Elimination half-life (T %) will be        calculated according to the following equation:

T _(1/2)=0.693/λ_(Z)

-   -   Area under the curve to the final sample with a concentration        greater than the LOQ (AUC_(last)) will be calculated using the        linear trapezoidal method and extrapolated to infinity using:

AUC _(inf) =AUC _(last) +C _(last)/λ_(Z)

-   -   where C_(last) is the final concentration LOQ.    -   All evaluable subjects completing at least one study period will        be included in the pharmacokinetic and statistical analysis.        Pharmacokinetic calculations will be performed using appropriate        software, e.g. WinNonlin (Pharsight Corporation) and/or SAS® for        Windows® (SAS Institute).    -   The relative bioavailability of the test formulation of naproxen        will be assessed under fasting and fed conditions using        AUC_(last) and AUC_(inf) after the 2×200 mg treatments        (Treatment A-fed, Treatment B-fasting), compared to the 1×500 mg        Naprosyn treatments (Treatment D-fed, Treatment E-fasting). The        relative bioavailability will be calculated for individual        subjects according to the following equation,

F=[Dose(ref)*AUC(test)]/[Dose(test)*AUC(ref)],

-   -   where Dose(ref)=500 mg, Dose(test)=400 mg, AUC(test)=AUC_(last)        or AUC_(inf) after administration of the test formulation, and        AUC(ref)=AUC_(last) or AUC_(inf) after administration of the        reference product. Fasting and fed treatments will be assessed        separately and the bioavailability estimates under each        condition will be summarized using descriptive statistics.    -   The dose-proportionality of naproxen in the test formulation        will be assessed using data acquired after administration of        Treatment B (2×200 mg, fasting) and Treatment C (1×200 mg,        fasting). The pharmacokinetic exposure parameters C_(max),        AUC_(last), and AUC_(inf) for individual subjects will be        dose-normalized by dividing through by the administered dose        (200 mg or 400 mg). The dose-normalized parameters will then be        compared using an ANOVA model, as described in Section 8.3.    -   Statistical Analysis    -   Comparison of the log-transformed pharmacokinetic parameters        C_(max), AUC_(last), and AUC_(inf) for naproxen across        treatments will be performed using an analysis of variance        (ANOVA) model and the two one-sided t-tests procedure. The ANOVA        model will include factors for sequence, subject within        sequence, treatment, and period. The ratios of the geometric        means (test to reference) and 90% confidence intervals will be        reported. Statistical analyses will be performed using        appropriate software, e.g. WinNonlin (Pharsight Corporation)        and/or SAS° for Windows® (SAS Institute).

Drug Supplies

Sufficient quantities of the study drug formulation to allow completionof this study will be supplied. Study drug formulations of naproxen 200mg capsules and Naprosyn® 500 mg tablets will be shipped to the clinicalresearch site pursuant to site Standard Operating Procedures (SOPs).Retention samples of investigational naproxen will not be required. Uponreceipt of the study drug products, the supplies will be inventoried andstored in an environmentally controlled and secure, limited access area.The lot numbers of the drugs along with the expiration dates (whereavailable) will be recorded and copies of the Certificate of Analysis(where available) will be maintained on file.

Records will be maintained of the receipt and dispensation of the drugssupplied. At the conclusion of the study, any unused study drug will bereturned to the sponsor or destroyed by the site pursuant to writtenauthorization by the sponsor and applicable federal and stateregulations.

Administrative Issues

The Investigator is referred to the Naprosyn® package insert,information provided during the study initiation visit, informationprovided by the study monitor, and ICH Guidelines for Good ClinicalPractice for information regarding the study drug, details, or generalconsiderations to be followed during the course of this study.

Events Schedule

SCREEN- END-OF- PROCEDURE ING STUDY STUDY Informed consent X Medical andmedication histories X X ECG X X Vital signs X X X Physical examinationX X Biochemistry, hematology, urinalysis X X Serology X Urine drugscreen X Urine drug and alcohol screen X Pregnancy test (femalesubjects) X X Standard high-fat, high-calorie X breakfast¹ Drugadministration X Blood sample collection for X pharmacokinetic analysisAdverse events X X ¹Treatments A and D only. Refer to protocol text fordetails.

Example 16

This Example describes a Phase 2, Randomized, Double-Blind, Single-Dose,Parallel-Group, Active- and Placebo-Controlled Study of NaproxenNanoformulation Capsules for the Treatment of Pain After SurgicalRemoval of Impacted Third Molars

The phase II efficacy study described in this example uses NaproxenNanoformulation Capsules 200 mg manufactured as described in Example 13.

Objectives:

The primary objective of this study is to evaluate the analgesicefficacy and safety of Naproxen Nanoformulation Capsules compared withplacebo in subjects with acute dental pain after third molar extraction.The secondary objective of this study is to evaluate the time to onsetof analgesia for Naproxen Nanoformulation Capsules compared with thestandard formulation of Naprosyn.

Number of Subjects:

Planned enrollment (and/or completion): Approximately 250 subjects (50in each treatment group) will be enrolled.

Subject Population:

Inclusion Criteria:

A subject will be eligible for study entry if all of the followinginclusion criteria are met:

-   -   1. Is male or female ≧18 and ≦50 years of age.    -   2. Requires extraction of 2 or more third molars. At least 1 of        the third molars must be a fully or partially bone-impacted        mandibular molar. If only 2 molars are removed, then they must        be ipsilateral.    -   3. Experiences moderate to severe pain intensity within 6 hours        after surgery, as measured by a Visual Analog Scale (VAS) score        of ≧50 mm on a 100-mm scale.    -   4. Has a body weight of ≧45 kg and a body mass index (BMI) ≦35        kg/m².    -   5. If female and of childbearing potential, is nonlactating and        nonpregnant (has negative pregnancy test results at screening        [serum] and on the day of surgery prior to surgery [urine]).    -   6. If female, is either not of childbearing potential (defined        as postmenopausal for at least 1 year or surgically sterile        [bilateral tubal ligation, bilateral oophorectomy, or        hysterectomy]) or practicing 1 of the following medically        acceptable methods of birth control:        -   a. Hormonal methods such as oral, implantable, injectable,            or transdermal contraceptives for a minimum of 1 full cycle            (based on the subject's usual menstrual cycle period) before            the study drug administration.        -   b. Total abstinence from sexual intercourse (since the last            menses before study drug administration).        -   c. Intrauterine device (IUD).        -   d. Double-barrier method (condoms sponge, diaphragm, or            vaginal ring with spermicidal jellies or cream).    -   7. Is in good health, in the opinion of the investigator.    -   8. Is able to provide written informed consent to participate in        the study and able to understand the procedures and study        requirements.    -   9. Must voluntarily sign and date an informed consent form (ICF)        that is approved by an Institutional Review Board (IRB) prior to        the conduct of any study procedure.    -   10. Is willing and able to comply with study requirements        (including diet and smoking restrictions), complete the pain        evaluations, remain at the study site overnight, and return for        follow-up 7±2 days after surgery.

Exclusion Criteria:

A subject will not be eligible for study entry if any of the followingexclusion criteria are met:

-   -   1. Has a known history of allergic reaction or clinically        significant intolerance to acetaminophen, aspirin, or any        nonsteroidal anti-inflammatory drug (NSAIDs, including        naproxen); history of NSAID-induced bronchospasm (subjects with        the triad of asthma, nasal polyps, and chronic rhinitis are at        greater risk for bronchospasm and should be considered        carefully); or hypersensitivity, allergy, or significant        reaction to sulfa (including sulfonamide) medicines, ingredients        of the study drug, or any other drugs used in the study        including anesthetics and antibiotics that may be required on        the day of surgery.    -   2. Has tested positive either on the urine drug screen or on the        alcohol breathalyzer test. Subjects who test positive at        screening only and can produce a prescription for the medication        from their physician may be considered for study enrollment at        the discretion of the investigator.    -   3. Has known or suspected history of alcoholism or drug abuse or        misuse within 2 years of screening or evidence of tolerance or        physical dependence before dosing with the study drug.    -   4. Has received or will require any medication (except hormonal        contraceptives, vitamins, or nutritional supplements) within 5        half-lives (or, if half-life is unknown, within 48 hours) before        dosing with study drug.    -   5. Has any clinically significant unstable cardiac, respiratory,        neurological, immunological, hematological, or renal disease or        any other condition that, in the opinion of the investigator,        could compromise the subject's welfare, ability to communicate        with the study staff, or otherwise contraindicate study        participation.    -   6. Has a history or current diagnosis of a significant        psychiatric disorder that, in the opinion of the investigator,        would affect the subject's ability to comply with the study        requirements.    -   7. Is receiving systemic chemotherapy, has an active malignancy        of any type, or has been diagnosed with cancer with 5 years of        screening (excluding squamous or basal cell carcinoma of the        skin).    -   8. Has a history of clinically significant (investigator        opinion) gastrointestinal (GI) event within 6 months before        screening or has any history of peptic or gastric ulcers or GI        bleeding.    -   9. Has a surgical or medical condition of the GI or renal system        that might significantly alter the absorption, distribution, or        excretion of any drug substance.    -   10. Is considered by the investigator, for any reason        (including, but not limited to, the risks described as        precautions, warnings, and contraindications in the current        version of the Investigator's Brochure [IB] for Naproxen        Nanoformulation Capsules), to be an unsuitable candidate to        receive the study drug.    -   11. Has history of chronic use (defined as daily use for >2        weeks) of NSAIDs, opiates, or glucocorticoids (except inhaled        nasal steroids and topical corticosteroids), for any condition        within 6 months before dosing with study drug. Aspirin at a        daily dose of 325 mg is allowed for cardiovascular (CV)        prophylaxis if the subject has been on a stable dose regimen for        30 days before screening and has not experienced any relevant        medical problem.    -   12. Has a significant renal or hepatic disease, as indicated by        the clinical laboratory assessment (results ≧3 times the upper        limit of normal [ULN] for any liver function test, including        aspartate aminotransferase [AST], alanine aminotransferase        [ALT], and lactate dehydrogenase, or creatinine ≧1.5 times the        ULN) or has any clinically significant laboratory findings at        screening that in the investigator's opinion contraindicate        study participation.    -   13. Has significant difficulties swallowing capsules or is        unable to tolerate oral medication.    -   14. Previously participated in another study of Naproxen        Nanoformulation Capsules, or received any investigational drug        or device or investigational therapy within 30 days before        screening.

Design:

This is a phase 2, multicenter, randomized, double-blind, single-dose,parallel-group, active- and placebo-controlled study to evaluate theefficacy and safety of Naproxen Nanoformulation Capsules (200 mg and 400mg doses) in subjects with postoperative dental pain. Eligible subjectswill complete all screening procedures within 28 days before thesurgery.

At screening, subjects will provide written informed consent toparticipate in the study before any protocol-specified procedures orassessments are completed. On Day 1, subjects who continue to beeligible for study participation after completing screening proceduresand assessments are will undergo extraction of 2 or more third molars.At least 1 of the third molars must be a fully or partiallybond-impacted mandibular molar. If only 2 molars are removed, then theymust be ipsilateral. All subjects will receive local anesthesia (2%lidocaine with 1:100,000 epinephrine). Nitrous oxide will be allowed atthe discretion of the investigator. Subjects who experience moderate tosevere pain intensity (a score of ≧50 mm on a 100-mm VAS) within 6 hoursafter surgery and who continue to meet all study entry criteria will berandomized in a 1:1:1:1:1 ratio to receive 1 oral dose of NaproxenNanoformulation Capsules (200 mg or 400 mg), Naprosyn tablets (250 mg or500 mg), or placebo. Study drug will be administered by an unblended,third-party doser who will not conduct any efficacy or safetyassessments.

Subjects will assess their baseline pain intensity (VAS) beforereceiving study drug (predose, Time 0) and their pain intensity (VAS)and pain relief (5-point categorical scale) at the following timepoints: 15, 30, and 45 minutes, and 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 10, and12 hours after Time 0; and immediately before the first dose of rescuemedication. The 2-stopwatch method will be used to record the time toperceptible and time to meaningful pain relief, respectively. Subjectswill complete a global evaluation of study drug 12 hours after Time 0 orimmediately before the first dose of rescue medication (whichever occursfirst). Vital signs will be recorded after the subject has been in asitting position for 5 minutes at the following times: before surgery,before Time 0, 12 hours after Time 0, and/or immediately before thefirst dose of rescue medication. Adverse events (AEs) will be monitoredand recorded from the time of signing of the ICF until the Follow-upVisit (or Early Termination Visit). During the 12 hours following Time0, subjects will complete efficacy and safety assessments. Subjects willremain at the study site overnight and will be discharged the morning ofDay 2. Upon discharge from the study site, subjects will be given adiary to record concomitant medications taken and AEs experienced afterdischarge.

Acetaminophen (1000 mg) will be permitted as the initial rescuemedication. Subjects will be encouraged to wait at least 60 minutesafter receiving study drug before taking rescue medication. Additionalanalgesic rescue medication may be administered at the discretion of theinvestigator if the protocol-specified rescue medication is deemedinadequate. Subjects are not permitted to take medications (excepthormonal contraceptives, vitamins, nutritional supplements, and studydrug) within 5 half-lives (or, if half-life is unknown, within 48 hours)before dosing with study drug until discharge from the study (Day 2).Other restrictions include the following: alcohol use is prohibited from24 hours before surgery until discharge on Day 2; nothing by mouth (NPO)from midnight before surgery until 1 hour after surgery; clear liquidsonly are allowed starting 1 hour after surgery until 1 hour afterdosing; 1 hour after dosing diet may be advanced according to standardpractice.

Upon discharge from the study site, subjects may be prescribed painmedication for use at home according to the standard practice of thestudy site. On Day 8 (±2 days), subjects will return to the study sitefor an abbreviated confirmatory physical assessment and concomitantmedication and AE assessments.

Study Drug:

Naproxen Nanoformulation Capsules (200 mg) for oral administration in asingle dose of either 200 mg (1 capsule) or 400 mg (2 capsules)

Reference Products:

Naprosyn tablets (250 mg and 500 mg)

Placebo capsule

Treatment Regimens

Eligible subjects meeting all study entry criteria will be randomized toreceive 1 of the following treatments:

Treatments Dose Naproxen 200 mg One 200-mg Naproxen NanoformulationCapsule Two placebo capsules Naproxen 400 mg Two 200-mg NaproxenNanoformulation Capsules One placebo capsule Naprosyn 250 mg One 250-mgNaprosyn tablet Two placebo capsules Naprosyn 500 mg One 500-mg Naprosyntablet Two placebo capsules Placebo Three placebo capsules

Study Duration:

Up to approximately 5 weeks for each subject, including a 4-weekscreening period and a posttreatment Follow-up Visit approximately 1week after dosing with study drug.

Investigative Sites or Countries:

Two study sites in the United States (US).

Study Endpoints:

Efficacy Endpoints:

The primary efficacy endpoint is the sum of total pain relief (TOTPAR)over 0 to 12 hours (TOTPAR-12) after Time 0.

The secondary endpoints are the following:

-   -   TOTPAR over 0 to 4 hours (TOTPAR-4) and over 0 to 8 hours        (TOTPAR-8) after Time 0.    -   VAS pain intensity difference (VASPID) at each scheduled time        point after Time 0.    -   Time to onset of analgesia (measured as time to perceptible pain        relief confirmed by meaningful pain relief).    -   VAS pain intensity score at each scheduled time point.    -   VAS summed pain intensity difference (VASSPID) over 0 to 4 hours        (VASSPID-4), over 0 to 8 hours (VASSPID-8), and over 0 to 12        hours (VASSPID-12) after Time 0.    -   Summed pain relief and intensity difference (sum of TOTPAR and        VASSPID [SPRID]) over 0 to 4 hours (SPRID-4), over 0 to 8 hours        (SPRID-8), and over 0 to 12 hours (SPRID-12) after Time 0.    -   Pain relief score at each scheduled time point after Time 0.    -   Peak pain relief.    -   Time to peak pain relief.    -   Time to first perceptible pain relief.    -   Time to meaningful pain relief.    -   Proportion of subjects using rescue medication.    -   Time to first use of rescue medication (duration of analgesia).    -   Patient's global evaluation of study drug.

Safety Endpoints:

The safety endpoints are the incidence of treatment-emergent AEs (TEAEs)and changes in vital sign measurements.

Statistical Methods Summary:

Analysis Populations:

The analysis populations include the following:

-   -   The intent-to-treat (ITT) population will consist of all        subjects who are treated with study drug and who have at least 1        pain relief assessment after Time 0. The ITT population is the        primary population for the efficacy analysis.    -   The per-protocol (PP) population will consist of all ITT        subjects who remain in the study for at least 12 hours of        treatment and who do not incur a major protocol violation that        would challenge the validity of their data. This population will        be utilized to evaluate the sensitivity of the primary efficacy        analysis.    -   The safety population will include all subjects who are treated        with study drug. The safety population is the population for all        safety assessments.

Subject Characteristics:

Demographic and baseline characteristics (including age, sec, race,weight, height, BMI, medical history, surgery duration, and baselinevalues of efficacy variables) will be summarized for each treatmentgroup and for the overall population by descriptive statistics. Noformal statistical analyses will be performed.

Efficacy Analyses:

The null hypothesis in this study is that TOTPAR-12 for placebo is equalto TOTPAR-12 for the 400-mg dose of Naproxen Nanoformulation Capsules.It will be analyzed using analysis of covariance (ANCOVA) models, whichinclude treatment effect and significant covariates. The effect ofpotential covariates, such as sex, baseline pain intensity, and surgicaltrauma rating, will be assessed using appropriate ANCOVA models. Theanalysis will be based on a 2-sided test as the significance level of0.05.

Other comparisons between the treatment regimens, including the 200-mgdose of Naproxen Nanoformulation Capsules versus placebo, 250-mgNaprosyn tablets versus placebo, and 500-mg Naprosyn tablets versusplacebo, will be considered secondary. No P value adjustment will bemade for multiple endpoints or multiple comparisons. Each efficacyendpoint will be summarized descriptively by treatment group.

For ordinal secondary endpoints, such as pain relief at each scheduledtime point, peak pain relief, and global evaluation of study drug,descriptive summaries will be provided to include the number andpercentage of subjects within each category for each treatment group.Nominal P values from Fisher's exact tests (or chi-square tests, asappropriate) comparing the placebo group with other treatment groupswill be provided, but no formal statistical inferences will be drawn onthe basis of these tests.

For each time-to-event endpoint, the Kaplan-Meier method will be used toevaluate the treatment effect. Time to onset of analgesia (measured astime to perceptible pain relief confirmed by meaningful pain relief)will be based on data collected using the 2-stopwatch method. Time toonset of analgesia will be right-censored at 12 hours for subjects whodo not experience both perceptible pain relief and meaningful painrelief during the 12-hour interval after Time 0. For time to onset ofanalgesia, the comparisons of interest will be the 200 mg NaproxenNanoformulation treatment group versus the 250 mg Naprosyn group and the400 mg Naproxen Nanoformulation treatment group versus the 500 mgNaprosyn group. The summary table will provide the number of subjectsanalyzed, the number of subjects censored, estimates for the quartiles,and 95% confidence intervals (Cis) for the estimated median and therestricted mean estimate. P values form the Wilcoxon or log-rank tests(as appropriate) will also be used to examine treatment effect. Coxproportional hazard models will be used to explore such potentialcovariates as sex, baseline pain intensity, and surgical trauma rating,if appropriate.

For the proportion of subjects using rescue medication, a logisticregression model that adjusts for baseline pain intensity, ifappropriate, will be used to evaluate the treatment effect. Subgroupanalysis by sex may be performed if it is confirmed to be astatistically significant covariate for TOTPAR-12. Baseline values aredefined as the last measurements taken before dosing with a study drug.

For pain intensity, missing observations will be imputed usingbaseline-observation-carried-forward (BOCF) for subjects who withdrawfrom the study due to lack of efficacy or an AE/intolerance to studydrug. The BOCF imputation will be applied in place of all scheduledassessments after the time of early termination due to lack of efficacyor an AE/intolerance to study drug using the baseline observation takenbefore Time 0.

For subjects who withdraw from the study due to reasons other than lackof efficacy or an AE/intolerance to study drug, missing observations forpain intensity and pain relief will be imputed usinglast-observation-carried-forward (LOCF). The LOCF imputation will beapplied in place of all scheduled assessments after the time of earlytermination due to reasons other than lack of efficacy or anAE/intolerance to the drug.

For subjects who take any dose of rescue medication, subsequent measuresafter the first dose of rescue medication will be disregarded. Instead,all scheduled assessments after the first dose of rescue medication willbe imputed using BOCF using the baseline observation taken before Time0. Single missing data points will be imputed using linearinterpolation, if they do not occur at the end of the study. For otherconditions before early termination or rescue medication, missing datawill be imputed using LOCF.

Safety Analysis:

Data listings will be provided for protocol-specified safety data. TheMedical Dictionary for Regulatory Activities (MedDRA) (Version 9.1 orhigher) will be used to classify all AEs with respect to system organclass and preferred term. Adverse event summaries will include onlyTEAEs, which will be summarized for each treatment group. Fisher's2-sided exact test will be used to compare the rates of occurrencebetween the placebo and Naproxen Nanoformulation Capsule groups for allTEAEs.

For vital sign measurements, descriptive statistics will be provided ateach scheduled time point for each treatment group. Changes fromBaseline for vital signs will be calculated for each subject, anddescriptive statistics will be provided on changes in vital signs fromBaseline for each treatment group at each scheduled time point afterBaseline. No formal statistical tests will be performed.

Sample Size:

The standard deviation of TOTPAR-12 is assumed to be ≦14.0. A samplesize of 50 subjects per treatment group will provide ≧80% power todetect a minimal difference of 8.0 in TOTPAR-12 using a 2-sample t-testwith a 0.05 two-sided significance level (nQuery v6.0).

TABLE 16a Schedule of Events B^(a) D A C E F G H I J K Written InformedConsent X Assign a screening number X Inclusion/exclusion criteria X XDemographics X Medical History X X^(b) Physical Examination^(c) X XVital signs^(d) X X X X X Height, weight, and BMI X Clinical laboratorytests (hematology, chemistry, X urinalysis) Pregnancy test for femalesubjects of X X childbearing potential^(c) Urine drug screen X X Alcoholbreathalyzer test X Oral radiography^(f) X Review study restrictionswith subject X Pain intensity (VAS)^(g) X X X X Randomization X Dosingwith study drug X Stopwatch assessment^(h) X Pain relief (5-pointcategorical scale)^(g) X X X Global evaluation of study drug^(i) XConcomitant medications X^(b) X X X X X X X Adverse events^(j) X X X X XX X X Dispense rescue medication/pain medications X Collect unusedrescue medication/pain X medications Dispense/collect subject diary X XDischarge from study site X A: Screening (Days −28 to −1); B: Day ofSurgery (Day 1); C: Preop; D: Postop; E: Predose; F: 0 h; G: 15, 30, 45min; H: 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 10 h; I: 12 h; J: Day 2; K:Follow-up (Day 8 ± 2 days or ET). Abbreviations: BMI, body mass index;ET, early termination; h, hour; min, minute; preop, preoperative;postop, postoperative; VAS, Visual Analogue Scale. ^(a)Times listed arerelative to dosing with study drug. ^(b)Medical history and concomitantmedication use since screening will be updated on Day 1 before surgery.^(c)A complete physical examination (excluding the genitourinaryexamination) will be performed at screening. An abbreviated confirmatoryphysical assessment, including an examination of the subject's mouth andneck, will be performed at the Follow-Up visit (or Early Terminationvisit) ^(d)Vital signs will be recorded after the subject has been in asitting position for 5 minutes at the following times: at screening,before surgery, before Time 0, 12 hours after Time 0, and/or immediatelybefore the first dose of rescue medication, and the Follow-up Visit (orEarly Termination visit). ^(e) Serum pregnancy test at screening andurine pregnancy test before surgery on Day 1 (female subjects ofchildbearing potential only). Test results must be negative for thesubject to continue in the study. ^(f)Oral radiographs taken within 1year before screening will be acceptable and do not need to be repeated.^(g)Pain assessments will be conducted at 15, 30, and 45 minutes and 1,1.5, 2, 3, 4, 5, 6, 7, 8, 10, and 12 hours after Time 0 and immediatelybefore the first dose of rescue medication. Pain intensity will also beassessed predose. At each assessment time point, the pain intensityassessment will be completed first and the pain relief assessment willbe completed second. Subjects will not be able to compare theirresponses with their previous responses. ^(h)Two stopwatches will bestarted immediately after the subject has swallowed the study drug with8 ounces of water (Time 0). Subjects will record the time to firstperceptible and meaningful pain relief, respectively, by stopping thestopwatches. ^(i)Subjects will complete a global evaluation of studydrug 12 hours after Time 0 or immediately before the first dose ofrescue medication (whichever occurs first). ^(j)Adverse events will bemonitored and recorded from the time of signing of the informed consentform (ICF) until the Follow-up Visit (or Early Termination visit).

Abbreviations: BMI, body mass index; ET, early termination; h, hour;min, minute; preop, preoperative; postop, postoperative; VAS, VisualAnalogue Scale.

-   -   a Times listed are relative to dosing with study drug.    -   b Medical history and concomitant medication use since screening        will be updated on Day 1 before surgery.    -   c A complete physical examination (excluding the genitourinary        examination) will be performed at screening. An abbreviated        confirmatory physical assessment, including an examination of        the subject's mouth and neck, will be performed at the Follow-Up        visit (or Early Termination visit)    -   d Vital signs will be recorded after the subject has been in a        sitting position for 5 minutes at the following times: at        screening, before surgery, before Time 0, 12 hours after Time 0,        and/or immediately before the first dose of rescue medication,        and the Follow-up Visit (or Early Termination visit).    -   e Serum pregnancy test at screening and urine pregnancy test        before surgery on Day 1 (female subjects of childbearing        potential only). Test results must be negative for the subject        to continue in the study.    -   f Oral radiographs taken within 1 year before screening will be        acceptable and do not need to be repeated.    -   g Pain assessments will be conducted at 15, 30, and 45 minutes        and 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 10, and 12 hours after Time 0        and immediately before the first dose of rescue medication. Pain        intensity will also be assessed predose. At each assessment time        point, the pain intensity assessment will be completed first and        the pain relief assessment will be completed second. Subjects        will not be able to compare their responses with their previous        responses.    -   h Two stopwatches will be started immediately after the subject        has swallowed the study drug with 8 ounces of water (Time 0).        Subjects will record the time to first perceptible and        meaningful pain relief, respectively, by stopping the        stopwatches.    -   i Subjects will complete a global evaluation of study drug 12        hours after Time 0 or immediately before the first dose of        rescue medication (whichever occurs first).    -   j Adverse events will be monitored and recorded from the time of        signing of the informed consent form (ICF) until the Follow-up        Visit (or Early Termination visit).

1. A method for producing a composition, comprising the steps of: dry milling a solid biologically active material and a millable grinding matrix in a mill comprising a plurality of milling bodies, for a time period sufficient to produce particles of the biologically active material dispersed in an at least partially milled grinding material wherein the biologically active material is naproxen. 